Open Access Opinion Article

The Double Helix in Retrospect: A Critique of Available Evidence

Sosale Chandrasekhar

International Journal of Biochemistry Research & Review, Page 1-6
DOI: 10.9734/IJBCRR/2016/31086

This short review commentary deals with the evidence for the DNA double helix and raises the intriguing question: does it exist to any significant extent in the solution phase? Well past the sixtieth year of its discovery via X-ray diffraction, the DNA double helix has now acquired the status of a sacred truth, with a large area of modern science revolving around it. This paper presents the pros and cons of the double helix from a physico-chemical perspective, although with critical inputs from chemical biology as appropriate.

Open Access Original Research Article

The Effect of Phorbol 12-Myristate 13-Acetate on CD11b and CD62-L Cell Surface Expression of Neutrophils and Monocytes

Jacob A. Mear, Stephen F. Hughes

International Journal of Biochemistry Research & Review, Page 1-12
DOI: 10.9734/IJBCRR/2016/30998

Background: Neutrophils and monocytes are phagocytic leukocytes that represent an important component of the non-specific immune system, by which invading micro-organisms and tumour cells are rapidly internalized and destroyed, via a number of biological mechanisms. Additionally, these phagocytic leukocytes play a key role in facilitating leukocyte-endothelial interactions in response to pro-inflammatory agents, during an inflammation. The expression of specific adhesion molecules such as CD62-L and CD11b, on the cell surface of neutrophils and monocytes, play an integral role with regards to cell migration, rolling, firm adhesion and subsequent cellular activation (leukocyte adhesion cascade). Leukocytes can be stimulated by chemotactic agents such as Phorbol 12-myristate 13-acetate (PMA). PMA is a direct stimulant of protein kinase C signaling pathway, which promotes cellular activation.

Methods: The aim of this pilot study was to determine the effect of PMA on CD62-L and CD11b cell surface expression of neutrophils and monocytes. Venous blood samples were collected from the ante-cubital fossa from healthy individuals, following written informed consent (n=4). Neutrophils and monocytes were isolated via a density gradient centrifugation method. Subsequently, neutrophil and monocyte cell surface expression of CD62-L and CD11b were measured by labelling with fluorescent anti-CD62-L and anti-CD11b monoclonal antibodies via flow cytometry.

Results: Following stimulation with PMA, monocytes displayed a decrease in CD62-L and increase in CD11b cell surface expression (p=0.103 and p=0.026 respectively). With respect to neutrophils, PMA resulted in a similar cellular response, where the cell surface expression of CD62-L decreased whilst CD11b increased (p=0.098 and p=0.025 respectively).

Discussion and Conclusion: In summary, this pilot study confirms the potent effect that PMA has on leukocyte function. Although studies involving PMA have previously been documented, this study provides a sound platform to continue work in this field of leukocyte biology. However, the validation and reliability of these findings would require to be assessed through the assessment of larger cohorts.

Open Access Original Research Article

Oxidation of Electron-rich Pollutants Using Peroxidase from Fenugreek Waste

Mugdha Ambatkar, Usha Mukundan, Himanshu Dawda

International Journal of Biochemistry Research & Review, Page 1-8
DOI: 10.9734/IJBCRR/2016/30500

Aims: To assess the capacity of peroxidase from non-edible fenugreek waste for removal of phenol, a model electron-rich pollutant, from synthetic wastewater.

Study Design: This study involved one-factor-at-a-time analysis. Reaction parameters such as pH of reaction mixture and concentrations of phenol and hydrogen peroxide were optimized one at a time.

Place and Duration of Study: Plant Biotechnology Research Laboratory, Ramniranjan Jhunjhunwala College, Mumbai between 2014 and 2015.

Methodology: The oxidative removal of the model pollutant, phenol, was assessed at ambient temperatures. The pH as well as concentrations of phenol and hydrogen peroxide were varied to optimize each reaction parameter. Each parameter was optimized singly while maintaining the other parameters constant.   

Results: The maximum activity (in guaiacol units) of peroxidase was observed at pH 6.0 in the presence of 6 mmol L−1 hydrogen peroxide. The removal of phenol achieved was maximum, 64.29% in 24 h, when the initial phenol concentration was 2 mmol L−1. Phenol removal proceeded through oxidative polymerization and settling of flocs.

Conclusion: The non-edible waste from fenugreek is a rich source of crude peroxidase from which the enzyme can be easily extracted. The use of vegetable waste as a source of a bioremediative enzyme is the primary focus of this work. This crude enzyme has potential for use in wastewater treatment by the oxidative degradation of electron-rich pollutants, such as phenols, amines, and azo dyes.

Open Access Original Research Article

Mechanistic Insight of Transient Receptor Potential Melastatin 8 Role in Streptozotocin Induced Diabetic Nephropathy

Doaa Hussein Zineldeen, Naglaa Ibrahim Sarhan, Nahid Mohamed Tahoon, Rania El-Sayed Wasfy

International Journal of Biochemistry Research & Review, Page 1-17
DOI: 10.9734/IJBCRR/2016/31435

Background: The transient receptor potential melastatin 8 (TRPM8) channel is a cold-sensing non-selective cation channel involved in cellular proliferation and signaling, yet its role in diabetic nephropathy (DN) remains poorly characterized. The TRPM8 agonist, geraniol (GE) is a dietary acyclic monoterpene alcohol known for its anti-oxidant, hypoglycemic and renal chemoprotective potentials. This study aims at defining the role of TRPM8 via the use of its agonist GE in an animal model of diabetic nephropathy.

Methods: A total of 80 male Wistar rats were equally divided into 4 equal groups: control, GE sham, diabetic rats received a single intraperitoneal injection of streptozotocin (STZ) (65 mg/kg) and STZ+GE diabetic rats received GE orally at a dose of (100 mg/kg/day). Indices of DNA damage, 8- Hydroxyguanosine (8-OHdG), nephrotoxicity and metabolic derangement parameters were measured eight weeks after diabetes induction. Real-time PCR was performed to detect mRNA expression levels of renal TRPM8 and podocyte marker. Renal histopathological and ultrastructural changes were recorded. Western blotting and immunohistochemistry were performed to determine TRPM8 protein expression.

Results: Diabetic rats displayed downregulation of TRPM8 mRNA and protein expression levels in renal tissues. Upon administration of GE, biochemical, ultrastructural and oxidative stress findings were significantly improved in treated diabetic animals compared to control groups and coincided with upregulation of renal TRPM8 expression as well as enhancement of podocytopathy.

Conclusion: The present study revealed that the ameliorative effect of GE in DN is TRPM8 mediated and highlights a mechanistic role of TRPM8 and its agonists in management of diabetic renal complications.

Open Access Original Research Article

Genetic Diversity and Relationships between and within Kiwifruit (Actinidia) Wild Species and Cultivated Varieties Using SRAP Markers

Zhao Bin Jing, Ming Xu, Yu Shan Lei

International Journal of Biochemistry Research & Review, Page 1-7
DOI: 10.9734/IJBCRR/2016/29091

Introduction: Kiwifruit (Actinidia) is one of the most important fruits in the world. Genetic diversity may provide the raw materials for programmers of plant breeding and crop improvement.

Materials and Methods: The aim of this study was to reveal the genetic diversity and relationships of 30 kiwifruit genotypes belonging to twelve different species using the SRAP marker.

Results: A total of 292 polymorphic bands were observed, with an average of 24.33 bands per pair of combined primers. The unweighted pair-group method of arithmetic average (UPGMA) analysis showed that the Jaccard’s coefficient of similarity value varied from 0.15 to 0.77, indicating that abundant diversities exist among these wild species. The 30 kiwifruit genotypes were divided into five groups using the cluster analysis and principal coordinate analysis. A. rufa and A. arguta had the far relationship, A. chinensis, A. deliciosa, and A. eriantha had the close genetic relationships.

Conclusion: This study provided theoretical basis for the genetic diversity and further breeding programs of Actinidia.