Assessing product adulteration of Eurycoma longifolia (Tongkat Ali) herbal medicinal product using DNA barcoding and HPLC analysis

Context: Eurycoma longifolia Jack (Simaroubaceae) commonly known as Tongkat Ali is one of the most important plants in Malaysia. The plant extracts (particularly roots) are widely used for the treatment of cough and fever besides having antimalarial, antidiabetic, anticancer and aphrodisiac activities.

Objectives: This study assesses the extent of adulteration of E. longifolia herbal medicinal products (HMPs) using DNA barcoding validated by HPLC analysis.

Materials and methods: Chloroplastic rbcL and nuclear ITS2 barcode regions were used in the present study. The sequences generated from E. longifolia HMPs were compared to sequences in the GenBank using MEGABLAST to verify their taxonomic identity. These results were verified by neighbor-joining tree analysis in which branches of unknown specimen are compared to the reference sequences established from this study and other retrieved from the GenBank. The HMPs were also analysed using HPLC analysis for the presence of eurycomanone bioactive marker.

Results: Identification using DNA barcoding revealed that 37% of the tested HMPs were authentic while 27% were adulterated with the ITS2 barcode region proven to be the ideal marker. The validation of the authenticity using HPLC analysis showed a situation in which a species which was identified as authentic was found not to contain the expected chemical compound.

Discussion and conclusions: DNA barcoding should be used as the first screening step for testing of HMPs raw materials. However, integration of DNA barcoding with HPLC analysis will help to provide detailed knowledge about the safety and efficacy of the HMPs.     

Comparison between self-reported physical activity (IPAQ-SF) and pedometer among overweight and obese women in the MyBFF@home study

Background

Several methods have been developed to determine a person’s physical activity level. However, there is limited evidence in determining whether someone is physically active or not. This study aims to determine the level of physical activity and to compare the usage of short version International Physical Activity Questionnaire (IPAQ-SF) and pedometer among overweight and obese women who were involved in the My Body is Fit and Fabulous at home (MyBFF@home) study.

Methods

Baseline and sixth month data from the MyBFF@home study were used for this purpose. A total of 169 of overweight and obese respondents answered the IPAQ-SF and were asked to use a pedometer for 7 days. Data from IPAQ-SF were categorised as inactive and active while data from pedometer were categorised as insufficiently active and sufficiently active by standard classification. Data on sociodemographic and anthropometry were also obtained. Cohen’s kappa was applied to measure the agreement of IPAQ-SF and pedometer in determining the physical activity level. Pre-post cross tabulation table was created to evaluate the changes in physical activity over 6 months.

Results

From 169 available respondents, 167 (98.8%) completed the IPAQ-SF and 107 (63.3%) utilised the pedometer. A total of 102 (61.1%) respondents were categorised as active from the IPAQ-SF. Meanwhile, only 9 (8.4%) respondents were categorised as sufficiently active via pedometer. Cohen’s κ found there was a poor agreement between the two methods, κ?=?0.055, p?>?0.05. After sixth months, there was +?9.4% increment in respondents who were active when assessed by IPAQ-SF but ??1.3% reductions for respondents being sufficiently active when assessed by pedometer. McNemar’s test determined that there was no significant difference in the proportion of inactive and active respondents by IPAQ-SF or sufficiently active and insufficiently active by pedometer from the baseline and sixth month of intervention.

Conclusion

The IPAQ-SF and pedometer were both able to measure physical activity. However, poor agreement between these two methods were observed among overweight and obese women.

     

Inflammation is more persistent in type 1 diabetic mice

Whether diabetes enhances or diminishes the host response to bacteria has been controversial. To determine how diabetes alters the inflammatory response, we inoculated P. gingivalis into the scalps of mice rendered diabetic with multiple low-dose streptozotocin treatment. On day 1, a moderate to severe inflammatory infiltrate was noted in both the diabetic and normoglycemic mice. After 3 days, the inflammatory infiltrate was significantly higher in the diabetic compared with the control group (P < 0.05). The mRNA expression of chemokines macrophage inflammatory protein-2 and monocyte chemoattractant protein-1 was strongly and similarly induced 3 hrs and 1 day post-inoculation. By day 3, the levels were reduced in normoglycemic mice but remained significantly higher in the diabetic group (P < 0.05). To determine whether persistent inflammation was specific for the streptozotocin-induced diabetic model, we directly compared the expression of TNF-alpha in streptozotocin-induced and db/db diabetic mice, which developed type 2 diabetes. Both exhibited prolonged TNF-alpha expression compared with controls. These results suggest that diabetes alters bacteria-host interactions by prolonging the inflammatory response.     

A systematic review of the applications of free tissue transfer in burns

Introduction

Free tissue transfer is a rarely indicated procedure in burns. However, in well selected cases it may play a pivotal role in optimizing outcomes in both primary and secondary burn reconstruction. We undertook a systematic review, based on the PRISMA statement for systematic reviews, of all published literature relating to the use of free flaps in acute burns and in secondary reconstructive procedures.

Methods

Inclusion and exclusion criteria were defined and Medline, Embase, PubMed and Google Scholar databases were searched from 1980 onwards to May 2013 with the search terms: “free flaps”, “free tissue transfer”, “microvascular”, “burns”, “acute burns”, “primary reconstruction” and “secondary reconstruction”.

Results

A total of 346 studies were retrieved following the search of which 30 studies met the inclusion criteria and were included in the review.

Discussion

We present the indications, timing, complications and failure rates for free flaps in primary and secondary reconstruction based on the available literature. We also provide a list of the various free flap options for the commonest sites undergoing reconstruction following burns. Finally an algorithm to ensure optimal success of free flaps when used in primary and in secondary burn reconstruction is presented.     

Reliability,Surgeon Preferences,and Eye-Tracking Assessment of the Stress Examination of the Ankle Syndesmosis

The diagnosis and stabilization of ankle syndesmotic injuries after acute injury remains an area of controversy in the foot and ankle surgical literature, seemingly without universal consensus. The primary objective of this investigation was to determine the frequency of agreement and reliability of the stress examination of the ankle syndesmosis. Secondary objectives were to determine surgeon preferences and protocols with respect to the ankle syndesmosis and to use gaze recognition software to perform an eye-tracking assessment during performance of stress examinations. Twelve foot and ankle surgeons, 12 residents, and 12 students were shown 5 intraoperative fluoroscopic still images and videos of the stress examination of the ankle syndesmosis. They were asked to evaluate the result as being “positive” or “negative” for syndesmotic stability. The overall reliability of the interpretation of the stress examination of the ankle syndesmosis was a kappa of 0.123 (surgeons 0.087; residents 0.019; students 0.237), indicating “slight” agreement. Survey results indicated wide variability in the perioperative preferences and protocols of surgeons dealing with the evaluation and treatment of the ankle syndesmosis. Eye-tracking results also demonstrated variability in the anatomic structures of interest focused on during performance of this testing. The results of this investigation provide evidence of reliability well below what would be expected of a gold standard test during stress examination of the ankle syndesmosis. These results indicate that future scientific endeavors are required to standardize the performance and interpretation of this testing.     

Association of C46T genetic polymorphism of coagulation factor XII with deep venous thrombosis: a cohort study on Egyptian patients

Deep vein thrombosis (DVT) is a common multi-factorial disease, with serious short- and long-term complications, and a potential fatal outcome. Many genes are involved in determining the interindividual variation in traits that define the onset and progression of disease, as well as the response to treatment. Several association studies have designed the relationship between factor XII C46T polymorphism and the risk of arterial and venous thrombosis. Some studies reported that FXII gene polymorphism is not associated with venous thrombosis, whereas other studies found an increased risk of venous thrombosis in carriers of a FXII-T variant. We constructed an age–gender–ethnic–matched case–control study including 52 DVT patients and 100 healthy volunteers. C46T polymorphism of the coagulation factor XII was carried out using allelic discrimination assay by real-time polymerase chain reaction for patients and controls, while plasma factor XII activity was detected by one-step clotting assay. FXII C46T genotyping in DVT patients revealed that 34.6% were heterozygous harboring the FXII-CT heterotype and 3.85% were homozygous; FXII-TT homotype, with no statistically significant difference in the distribution of the mutant genotypes between DVT patients and the control group. FXII activity was significantly reduced in DVT patients harboring the mutant genotypes. In the present study, FXII C46T gene polymorphism was not associated with increased risk of deep venous thrombosis.