Methods for detection and quantification of gelatin from different sources.

Gelatin is a water-soluble protein obtained from the collagen of various animal origins (porcine, bovine, fish, donkey, horse, and deer hide) and has diverse applications in the food, pharmaceutical, and cosmetics industries. Porcine and bovine gelatins are extensively used in food and non-food products; however, their acceptance is limited due to religious prohibitions, whereas fish gelatin is accepted in all religions. In Southeast Asia, especially in China, gelatin obtained from donkey and deer skins is used in medicines. However, both sources suffer from adulteration (mixing different sources of gelatin) due to their limited availability and high cost. Unclear labeling and limited information about actual gelatin sources in gelatin-containing products cause serious concern among societies for halal and fraud authentication of gelatin sources. Therefore, authenticating gelatin sources in gelatin-based products is challenging due to close similarities between the composition differences and degradation of DNA and protein biomarkers in processed gelatin. Thus, different methods have been proposed to identify and quantify different gelatin sources in pharmaceutical and food products. To the best of our knowledge, this systematic and comprehensive review highlights different authentication techniques and their limitations in gelatin detection and quantification in various commercial products. This review also describes halal authentication and adulteration prevention strategies of various gelatin sources, mainly focussing on research gaps, challenges, and future directions in this research area.

A rapid colorimetric method for the detection of carminic acid in samples based on visible color change.

Carminic Acid (CA), an insect-derived red color, is widely used as a colorant and additive in food and non-food items. The detection of CA is of great concern since it is unacceptable for vegetarians and vegans consumers. Therefore, it is important for food authorities to have a rapid detection method for CA. We describe here a simple and rapid method for the qualitative detection of CA, using Pb2+ for complex formation. As a result, the sample solution shows a visible change from pink to purple (bathochromic shift) which could also be analyzed through a spectrophotometer at λmax = 605 nm. The structure of the CA-Pb2+ complex was also studied through advanced spectroscopic techniques. Moreover, the presence of iron results in the formation of a stable CA-Fe2+ complex without any significant color change, as Fe2+ has a stronger binding affinity with CA. Thus, sodium fluoride (NaF) was used to prevent CA-Fe2+ complex formation. Therefore, two methods were developed based on the absence (method I) and presence (method II) of NaF. The LOD and LOQ for the method I was 0.0025 and 0.0076 mg mL-1, and for method II, values were 0.0136 and 0.0415 mg mL-1, respectively. The methods were also validated by intra and inter-day analyses. A total of 45 commercials, including food and non-food samples, were screened for the detection of CA. The developed methods are applicable for the effective and rapid surveillance of CA in various samples without the use of high-tech instruments.

Acyclovir induces fetal hemoglobin via downregulation of γ-globin repressors, BCL11A and SOX6 trans-acting factors.

Pharmacological reactivation of developmentally silenced fetal hemoglobin (HbF) is an attractive approach to ameliorate the clinical manifestations of ß-thalassemia and sickle cell anemia. Hydroxyurea, the only HbF inducer, has obtained regulatory approval. However, hydroxyurea non-responders and associated myelosuppression making its widespread use undesirable. A high level of HbF with safe and effective agents remains an elusive therapeutic goal for this global health burden. This study demonstrated the effect of acyclovir on γ-globin expression and erythropoiesis, associated with increased HbF production. In vitro, human erythroleukemia cells and human CD34+ erythroid progenitors, and in vivo ß-YAC transgenic mice were used as experimental models. We found that acyclovir significantly induces expression of the γ-globin gene and HbF synthesis in CD34+ erythroid progenitors, without affecting terminal erythroid differentiation and erythroid cell proliferation. In contrast to other HbF inducers, no associated cytotoxicity with acyclovir was observed. Further, we reported the effect of acyclovir on γ-globin gene transcriptional regulators including BCL11A, FOP1, KLF1 SOX6, and GATA-1. Significant downregulation of the γ-globin repressors BCL11A and SOX6 was observed at both mRNA and protein levels. Whereas, GATA-1, a master erythroid transcription factor, was upregulated in acyclovir treated human CD34+ erythroid culture. Similarly, the HbF inducing effect of acyclovir in ß-YAC transgenic mice revealed a good in vitro correlation, with a substantial increase in fetal globin mRNA, and F cells population. These findings collectively suggest acyclovir as an effective HbF inducer and pave the way to evaluate its clinical efficacy in treating ß-globin disorders.

Author Correction: Association of Cyclin Dependent Kinase 10 and Transcription Factor 2 during Human Corneal Epithelial Wound Healing in vitro model.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Association of Cyclin Dependent Kinase 10 and Transcription Factor 2 during Human Corneal Epithelial Wound Healing in vitro model.

Proper wound healing is dynamic in order to maintain the corneal integrity and transparency. Impaired or delayed corneal epithelial wound healing is one of the most frequently observed ocular defect and difficult to treat. Cyclin dependen kinase (cdk), a known cell cycle regulator, required for proper proliferating and migration of cell. We therefore investigated the role of cell cycle regulator cdk10, member of cdk family and its functional association with transcriptional factor (ETS2) at active phase of corneal epithelial cell migration. Our data showed that cdk10 was associated with ETS2, while its expression was upregulated at the active phase (18 hours) of cell migration and gradually decrease as the wound was completely closed. Topical treatment with anti-cdk10 and ETS2 antibodies delayed the wound closure time at higest concentration (10 µg/ml) compared to control. Further, our results also showed increased mRNA expression of cdk10 and ETS2 at active phase of migration at approximately 2 fold. Collectively, our data reveals that cdk10 and ETS2 efficiently involved during corneal wound healing. Further studies are warranted to better understand the mechanism and safety of topical cdk10 and ETS2 proteins in corneal epithelial wound-healing and its potential role for human disease treatment.

Electrospray tandem mass spectrometric analysis of a dimeric conjugate, salvialeriafone and related compounds.

BACKGROUND: Electrospray tandem mass spectrometry approach is widely used for the rapid characterization of natural products. This paper describes the gas-phased ESI-MS/MS fragmentation of abietane-type diterpenoids and their novel dimeric conjugate, salvialeriafone (1) using both positive and negative ion electrospray ionization quadropole time-of-flight mass spectrometry (ESI-QqTOF-MS/MS) hybrid instrument. Diterpenoids are widely distributed throughout the plant kingdom and posses interesting biological activities. RESULTS: ESI-QqTOF-MS (positive ion mode) of diterpenoids 1-6 under collision-induced dissociation tandem mass spectrometric analysis (CID-MS/MS) showed the characteristic losses of water, carbonmonoxide and propene molecules, while analysis in negative ion mode showed the characteristic losses of water, carbon monoxide, methane molecules and methyl radical. Results demonstrated the differences in the product ions and base peaks due to the differences in the skeleton. A novel dimeric conjugate, salvialeriafone (1) showed characteristic fragmentation pattern and was found to be more prone to form radical ions, as compared to monomeric diterpenoids. The fragmentation pathways of characteristic fragments were proposed with the aid of HRESIMS. CONCLUSIONS: Extensive tandem mass spectrometric studies of salvialeriafone (1) and related diterpenoids 2-6 were conducted and their characteristic fragments were identified. The knowledge of the fragmentation pattern of these diterpenoids will be useful for the characterization of new dimers of this class of compounds.