News Update on Vitamins: September 2021

 

Effect of irradiation on vitamins

Food irradiation is a physical process involving treatment of food with ionising radiation. Its main uses are reduction in spoilage and pathogenic organisms, inhibition of ripening and sprouting processes, and insect disinfestation. Chemical changes in the treated foods are small, and expert committees have concluded that they carry no special nutritional problems. Some vitamins are sensitive to irradiative degradation, however, and opponents of the process have claimed that extensive destruction will occur. Irradiation doses will, however, be limited by organoleptic changes, and maximum levels are being introduced into legislation for specific foods. Examination of the published literature shows that vitamins C and B1 are the most sensitive water-soluble vitamins, and that E and A are the most sensitive fat-soluble vitamins. Vitamin losses on irradiation of permitted foods in western countries will not be of nutritional importance. [1]

Chapter 59 – Hydrosoluble vitamins

The hydrosoluble vitamins are a group of organic substances that are required by humans in small amounts to prevent disorders of metabolism. Significant progress has been made in our understanding of the biochemical, physiologic and nutritional aspects of the water-soluble vitamins. Deficiency of these particular vitamins, most commonly due to inadequate nutrition, can result in disorders of the nervous system. Many of these disorders have been successfully prevented in developed countries; however, they are still common in developing countries. Of the hydrosoluble vitamins, the nervous system depends the most on vitamins B and C (ascorbic acid) for proper functioning. The B group vitamins include thiamin (vitamin B1), riboflavin (vitamin B2), niacin or niacinamide (vitamin B3), pantothenic acid (vitamin B5), pyridoxine or pyridoxal (vitamin B6) and cobalamin (vitamin B12). Clinical findings depend upon the deficiency of the underlying vitamin; generally, deficiency symptoms are seen from a combination rather than an isolated vitamin deficiency. True hereditary metabolic disorders and serious deficiency-associated diseases are rare and in general limited to particular geographic regions and high-risk groups. Their recognition is truly important as that determines the appropriate therapeutic management. The general availability of vitamins to practically everyone and several national health programs have saved many lives and prevented complications. However, there has been some apprehension for several decades about how harmless generous dosages of these vitamins are. Overt overdosages can cause vitamin toxicity affecting various body systems including the nervous system. Systemically, vitamin toxicity is associated with nonspecific symptoms, such as nausea, vomiting, diarrhea, and skin rash which are common with any acute or chronic vitamin overdose. At a national level, recommended daily allowances for vitamins become policy statements. Nutrition policy has far reaching implications in the food industry, in agriculture, and in food provision programs. Overall, water-soluble vitamins are complex molecular structures and even today, many areas of vitamin biochemistry still need to be explored. Many readers might be of the opinion that the classic forms of nutritional deficiency diseases have faded into the background of interesting history. This has caused their diverse symptoms to be neglected by most modern physicians since vitamin enrichment of many foods automatically erases them from their consideration in differential diagnosis. Vitamin B12 and folic acid deficiencies are discussed in other chapters. [2]

Vitamins and cancer

The prospect that high intake of certain vitamins may confer protection against cancer has drawn substantial attention during the last decades. This paper gives a concise update of the role of a number of promising vitamins in prevention of cancer. Vitamin A and its analogues have an important role in cellular processes related to carcinogenesis. However, blood vitamin A levels are under strict control and a high intake of preformed vitamin A does not seem to be relevant for cancer prevention. The antioxidant vitamins C and E and β-carotene may also have other biological activities than free radical trapping that relate to their cancer preventive properties. Mechanisms include immune stimulation, inhibition of nitrosamine formation, enhancement of cell communication and an influence on metabolic activation of carcinogens. Epidemiological data for the antioxidant vitamins are promising, but cannot rule out that another factor or combination of factors in fruits and vegetables might be responsible for a protective effect. The B vitamin folic acid is one of these potential factors that is currently thought to have an influence on DNA methylation and thus on proto-oncogene expression. Folic acid seems to be promising and deserves further study. Vitamin D might be relevant in colon cancer development due to its close links with calcium metabolism that might influence cell proliferation. Overall, results are promising, but the first human intervention trials on (antioxidant) vitamins and human cancer have yielded somewhat disappointing results. At this moment the data seem insufficient to make recommendations for vitamin supplementation to prevent cancer. The results are certainly in line with the advice that a diet rich in fruits and vegetables will help reduce cancer risk. [3]

Antihypercholesterolemic, Cardioprotective and Vitamins E and C Sparing Properties of Bryophyllum pinnatum in Rabbits

Potential of Bryophyllum pinnatum in preventing excessive lipaemia and oxidation, hence reducing predisposition to chronic diseases was assessed. B. pinnatum supplement was prepared by macerating a known weight of the plant in a known volume of distilled water. Ten rabbits were used for the study and were equally grouped into 2. Rabbits in group 1 were given 0.1 ml of distilled water once daily for 60 days and they served as control animals. A dose of 0.1 ml containing 0.2 g/ml of B. pinnatum supplement was given to each of the rabbits in group 2 once daily for 60 days. Bryophyllum pinnatum caused reduction in cholesterol concentration in serum, kidney, heart, intestine, brain and liver, while increased total lipids concentrations were observed in serum and kidney. B. pinnatum supplement reduced lipid peroxidation in heart, brain and liver. Administration of B. pinnatum caused increased glutathione reductase activity as shown by increased GSH concentration. Bryophyllum pinnatum demonstrated vitamins C and E sparing effects. Conclusively, B. pinnatum may be a good candidate as a nutraceutical. [4]

Influence of Packaging Materials and Storage Conditions on the Vitamins A and E Storage Stability of Palm Oil in Nigeria

Aims: Despite the presence of natural antioxidants contained in palm oil, it is still susceptible to quality deteriorations if not properly stored. This study therefore evaluates the storage stability of vitamins A and E in palm oil in four prominent packaging materials (metal cans, white plastic bottles, glass bottles and pet bottles) used in Nigeria and under three storage conditions [(refrigeration (5ºC), closed cupboard (27ºC) and direct sunlight (35ºC)].
Study Design: Freshly produced palm oil was filled in metal cans, white plastic bottles, glass bottles and pet bottles and stored in open, direct sunlight (35±1ºC), closed wooden cupboard (27±1ºC) and a refrigerator (5±1ºC) for a period of 120 days The samples were stored in a 4 (packaging materials) x 3 (Temperature) factorial arrangement making 12 treatments for each analysis sampled every 30 days for a period of 120 days. Vitamins A and E contents of palm oil samples were determined at 30 days intervals using ultraviolet spectrometer and high Performance Liquid Chromatography, respectively. Data values of triplicate determinations of vitamins A and E contents obtained from analysis were subjected to analysis of variance (ANOVA) and mean values were separated using Duncan New Multiple Range (DNMR) test using the Statistical Package for Social Sciences (SPSS) version 17.0. The rates of changes in the Vitamins A and E contents over the storage period of 120 days were also determined using Linear Regression analysis.
Place and Duration of Study: Department of Food Science and Technology, Federal University of Technology, Akure, Nigeria between January, 2012 and December 2013.
Methodology: Palm oil filled into the four different packaging materials was stored in the three storage conditions for a period of 120 days. Vitamins A and E contents of palm oil samples were determined at 30 days intervals using ultraviolet spectrometer and high Performance Liquid Chromatography, respectively. Data obtained were subjected to Analysis of Variance (ANOVA) to determine the statistical significant differences in the packaging materials and the storage conditions and the interactions between them. Mean values of vitamin A and vitamin E of different packaging methods and storage conditions were separated by Duncan New Multiple Range (DNMR) test to indicate their levels of significant differences. Linear Regression Analysis was also performed to determine the rates of changes in the vitamin A and E with time during storage.
Results: The vitamins A and E content of the samples stored in open, direct sunlight were virtually lost at the end of the storage period. For samples stored in sunlight, the vitamin A values in metal cans decreased by 97.45%, in white plastic bottle by 92.19%, in glass bottle by 92.46% and in pet bottle by 93.13% while vitamins E also decreased by 92.31%, 61.54%, 75.48% and 82.05%, respectively. Samples stored at room temperature suffered a higher amount of losses compared to the refrigerated samples. The refrigerated samples recorded only a minimal amount of loss. For the storage in both the sunlight and the dark cupboard and storage under refrigerating temperature of 5ºC, the order of preference for the packaging materials was white plastic bottle > glass bottle > pet bottle > metal can.
Conclusion: The results obtained from this study have demonstrated that packaging palm oil in white plastic bottle is the best method of preserving palm oil under refrigerating condition and lacquered metal under sunlight and dark cupboard. It has also shown that vitamins A and E degrade faster when palm oil is stored under sunlight and totally unfit for human consumption at the end of the storage period hence, palm oil should be stored in cold, dry places to limit their losses of antioxidant components. [5]

Reference

[1] Kilcast, D., 1994. Effect of irradiation on vitamins. Food chemistry, 49(2), pp.157-164.

[2] Chawla, J. and Kvarnberg, D., 2014. Hydrosoluble vitamins. Handbook of clinical neurology, 120, pp.891-914.

[3] van Poppel, G. and van den Berg, H., 1997. Vitamins and cancer. Cancer letters, 114(1-2), pp.195-202.

[4] Adekunle, A.S., Adelusi, T.I., Oyewo, E.B., Kamdem, J.P. and Akintade, B.B., 2016. Antihypercholesterolemic, cardioprotective and vitamins E and C sparing properties of Bryophyllum pinnatum in rabbits. European Journal of Medicinal Plants, pp.1-13.

[5] Oluwalana, I.B., Oluwamukomi, M.O., Toriola, B.O. and Karim, O.R., 2015. Influence of packaging materials and storage conditions on the vitamins A and E storage stability of palm oil in Nigeria. Advances in Research, pp.191-202.

Characterization of Polyhydroxyalkaonoate Extracted from Bacillus megaterium JHA and Its Biodegradation Studies

Polyhydroxyalkanoates (PHAs) are naturally occurring polymers that are generated as a source of energy by a variety of bacterial organisms. They can be used to replace petroleum-based plastics in a sustainable way. Bacillus megaterium JHA, a gram-positive bacterium isolated from oil-contaminated soil, demonstrated the ability to accumulate high levels of PHA on glucose as a substrate. The biopolymer was extracted from the above strain using a solvent extraction method (chloroform) to obtain a thin film of PHA in the current sample. Analytical techniques such as confocal microscopy, high performance thin layer chromatography, fourier transform infrared spectroscopy, nuclear magnetic resonance, and gel permeation chromatography were used to characterise this film. These techniques revealed a functional and chemical similarity between PHA and the standard molecule, Polyhydroxybutryrate (PHB), suggesting that it is a PHB derivative. A differential scanning calorimeter and thermo-gravimetric analysis were used to assess the biopolymer’s thermal properties. With a polydispersity index of 1.7, the biopolymer’s weight-average molecular weight and number-average molecular mass were determined to be 43.47kDa and 25.53kDa, respectively. It also revealed a melting temperature of 163.19°C, as well as a thermal decomposition temperature of 285.68°C. The biopolymer’s IC50 value was measured as 0.311 mg/mL using the MTT assay, indicating that it is suitable for a variety of biomedical applications. Scanning electron microscopy was used to further investigate the biopolymer’s biodegradability. Apart from the analytical characteristics listed above, the polymer’s unique attribute was its ability to degrade fully in compost soil under facultative conditions in 60-90 days.

Author(s) Details

Joyline Mascarenhas
Department of Microbiology, Wilson College, Mumbai 400007, Maharashtra, India.

K. Aruna
Department of Microbiology, Wilson College, Mumbai 400007, Maharashtra, India.

View Book:- https://stm.bookpi.org/CPEBJIBS/issue/view/24

Latest News on Acute Toxicity: Jan 2020

Measurement of pollutant toxicity to fish I. Bioassay methods for acute toxicity

The review describes profitable methods for measuring lethal levels of pollutants for aquatic organisms. Methods for research in the laboratory are emphasized but the same principles could be applied in field work. Greater use of standard toxicological methods and terminology is urged.

For 211 out of 375 toxicity tests reviewed, acute lethal action apparently ceased within 4 days, although this tabulation may have been biased towards short times by a large number of static tests.

The incipient LC50 (lethal concentration for 50 per cent of individuals on long exposure) is recommended as the most useful single criterion of toxicity. If this cannot be estimated, the 4-day LC50 is a useful substitute, and often its equivalent.

A desirable first step in toxicity tests is to estimate median lethal time for each of a series of concentrations. A toxicity curve should be drawn by plotting median survival times against concentrations on logarithmic paper. The curve helps to reveal any unusual features of toxicity. Whenever possible, tests should be prolonged until the toxicity curve becomes parallel to the time axis, indicating a lethal threshold concentration. The incipient LC50 is then estimated by selecting an exposure time from the asymptotic part of the toxicity curve; for this exposure time, observed mortality is plotted against concentration on log-probit paper, and the LC50 is read from an eye-fitted line.
Confidence limits of the LC50 may also be estimated by simplified methods. These should be given in published work along with a value for slope of the probit line. [1]

Acute toxicity of pesticides

LD50 values have been determined for 98 pesticides and 2 metabolites of DDT administered in a single dose by the oral or dermal route to Sherman strain adult rats. Most compounds tested by the oral route were more toxic to female than to male rats, but 9 of 85 compounds (aldrin, chlordane, heptachlor, Abate, Imidan, methyl parathion, ronnel, schradan, and metepa) tested in both sexes were markedly more toxic in male rats. Although the pesticides were generally more toxic by the oral than by the dermal route, 5 compounds (fenitrothion, Merphos, mevinphos, Isolan, and Omite) were more toxic by the dermal route. LD50 values for rats treated by the dermal route with parathion or dichlorvos and restrained so as to prevent the possibility of oral or respiratory exposure were not significantly different from those values in rats treated according to the procedure used routinely in this laboratory.

In screening tests for the production of paralytic effect in chicken hens 3 of 9 carbamate and 22 of 30 organic phosphorus pesticides tested caused paralysis. The onset of paralysis was delayed at least 3 days in chickens treated with Dursban and Merphos and at least 14 days in chickens treated with DEF. Chickens treated with the other 22 pesticides that produced paralysis showed this effect within 24 hours. [2]

Acute toxicity of ropivacaine compared with that of bupivacaine.

The acute central nervous and cardiovascular effects of the local anesthetics ropivacaine and bupivacaine were compared in 12 volunteers in a randomized double-blind manner with use of intravenous infusions at a rate of 10 mg/min up to a maximal dose of 150 mg. The volunteers were all healthy men. They were familiarized with the central nervous system (CNS) toxic effects of local anesthetics by receiving a preliminary intravenous injection of lidocaine. The infusions of ropivacaine and bupivacaine were given not less than 7 days apart. CNS toxicity was identified by the CNS symptoms and the volunteers were told to request that the infusion be stopped when they felt definite but not severe symptoms of toxicity such as numbness of the mouth, lightheadedness, and tinnitus. In the absence of definite symptoms, the infusion was stopped after 150 mg had been given. Cardiovascular system (CVS) changes in conductivity and myocardial contractility were monitored using an interpretive electrocardiograph (which measured PR interval, QRS duration, and QT interval corrected for heart rate) and echocardiography (which measured left ventricular dimensions from which stroke volume and ejection fraction were calculated). Ropivacaine caused less CNS symptoms and was at least 25% less toxic than bupivacaine in regard to the dose tolerated. Both drugs increased heart rate and arterial pressure. Stroke volume and ejection fraction were reduced. There was no change in cardiac output. Although both drugs caused evidence of depression of conductivity and contractility, these appeared at lower dosage and lower plasma concentrations with bupivacaine than with ropivacaine. [3]


In vitro Antisickling and Radical Scavenging Activities of a Poly-herbal Formula (Drepanoalpha®) in Sickle Cell Erythrocyteand Acute Toxicity Study in Wistar Albino Rats

Aims: To evaluate the antisickling and radical scavenging activities and acute toxicity of indigenous nutritive formula Drepanoalpha®, produced through a bio-guided based plant selection.

Study Design: Drepanoalpha® extracts, Antisickling activity by Emmel test, Antioxidant activity by 1,1-diphenyl-2-picrylhydrazyl bleaching methods; acute toxicity on rats, determination of biological and haematological parameters.

Place and Duration of Study: Science Faculty University of Kinshasa, between January 2013 and February 2014.

Methodology: The antisickling and antioxidant activities of Drepanoalpha® were determined using Emmel and the 1,1-diphenyl-2-picrylhydrazyl bleaching methods respectively. Acute oral toxicity test was performed to determine the LD50. Liver and kidney functions, the hematological and histopathological examinations were assessed using standard techniques.

Results: Obtained results revealed that Drepanoalpha® possessesinteresting in vitro antisickling and antioxidant activities as revealed by the observed normal biconcave form of sickle erythrocyte (normalization rate >80%) and the radical scavenging activity (ED50= 0.604 ± 0.028 µg/mL). Acute toxicity assessment revealed that the medium lethal dose (LD50) is higher than 4000 mg/kg. Drepanoalpha® significantly increases the values of WBC, RBC, Hb, HCT, PLT, IDR-CV and PCT. Furthermore, this polyherbal formula significantly decreases the values of IDR-SD, P-RGC, AST and ALT (p<0.05). Both the control and treated groups displayed comparable non altered histological architecture of the liver cells.

Discussion: The mean values of biochemical markers and hematological markers of treated rats revealed that Drépanoalpha® is potentially safe indicating non-toxic effect of the phytomedicine on immune cells and blood clotting factors. Moreover, this poly-herbal formulation increases the hemoglobin rate in the all treated rats (500-4000 mg/kg bodyweight) and preserves the histological architecture of the liver cells.

Conclusion: Drepanoalpha® may increase weight gain, promote erythropoiesis and thrombopoeisis in sicklers patients. This phytomedicine could be used in the treatment of all form of anemia and may also prevent bile duct obstruction or intra-hepatic cholestasis. The results can form the basis for clinical trials in humans. [4]

Acute Toxicity and Hypoglycemic Activity of Aqueous Fruit Pulp Extract of Adansonia digitata L. (Afpead) on Alloxan Induced Diabetic Rats

Oral LD50 and hypoglycemic activity of aqueous fruit pulp extract of Adansonia digitata L. (AFPEAD) were investigated in this research. A total of fourty eight (48) rats were used, twelve (12) of the rats were used for oral LD50 determination, and were grouped into four (4) groups of three rats (3) each. The first three groups were administered with 10 mg/kg, 100 mg/kg and 1000 mg/kg body weight of the extract respectively, while the last group was subdivided into three groups of one rat each and were administered with 1600 mg/kg, 2900 mg/kg and 5000 mg/kg body weight of the extract respectively. Thirty (36) rats were used for the diabetic study and were grouped into six (6) groups of six (6) rats each. Group I served as normal control, group II served as diabetic control while Groups III, IV, V and VI were induced with diabetes and administered with AFPEAD at a dose of 1.00 g/kg, 2.00 g/kg and 3.00 g/kg body weight and standard drug (Chlorpropamide, 100 mg/kg) respectively for two weeks. The research found the oral LD50 of the extract to be greater than 5000mg/kg indicating that the extract was practically non-toxic and administration of the extract to test groups shows a significant (p<0.05) decrease in blood glucose level when compared to diabetic control after two weeks treatment with the extract. Thus indicating a hypoglycemic activity by the extract which might be due to the presence of various phytochemicals. [5]

Reference

[1] Sprague, J.B., 1969. Measurement of pollutant toxicity to fish I. Bioassay methods for acute toxicity. Water research, 3(11), pp.793-821.

[2] Gaines, T.B., 1969. Acute toxicity of pesticides. Toxicology and applied pharmacology, 14(3), pp.515-534.

[3] Scott, D.B., Lee, A., Fagan, D., Bowler, G.M., Bloomfield, P. and Lundh, R., 1989. Acute toxicity of ropivacaine compared with that of bupivacaine. Anesthesia and analgesia, 69(5), pp.563-569.

[4] N. Ngbolua, K., T. Mpiana, P., S. T. Tshibangu, D., P. Mazasa, P., Z. Gbolo, B., K. Atibu, E., N. Kadima, J. and M. Kasali, F. (2014) “In vitro Antisickling and Radical Scavenging Activities of a Poly-herbal Formula (Drepanoalpha®) in Sickle Cell Erythrocyteand Acute Toxicity Study in Wistar Albino Rats”, European Journal of Medicinal Plants, 4(10), pp. 1251-1267. doi: 10.9734/EJMP/2014/11861.

[5] U. Muhammad, I., K. Jarumi, I., J. Alhassan, A., M. Wudil, A. and A. Dangambo, M. (2016) “Acute Toxicity and Hypoglycemic Activity of Aqueous Fruit Pulp Extract of Adansonia digitata L. (Afpead) on Alloxan Induced Diabetic Rats”, Journal of Advances in Medical and Pharmaceutical Sciences, 6(3), pp. 1-6. doi: 10.9734/JAMPS/2016/23862.

Latest News on RNA Isolation: Dec 2020

The single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction: twenty-something years on

Since its introduction, the ‘single-step’ method has become widely used for isolating total RNA from biological samples of different sources. The principle at the basis of the method is that RNA is separated from DNA after extraction with an acidic solution containing guanidinium thiocyanate, sodium acetate, phenol and chloroform, followed by centrifugation. Under acidic conditions, total RNA remains in the upper aqueous phase, while most of DNA and proteins remain either in the interphase or in the lower organic phase. Total RNA is then recovered by precipitation with isopropanol and can be used for several applications. The original protocol, enabling the isolation of RNA from cells and tissues in less than 4 hours, greatly advanced the analysis of gene expression in plant and animal models as well as in pathological samples, as demonstrated by the overwhelming number of citations the paper gained over 20 years. [1]

Importance of RNA isolation methods for analysis of exosomal RNA: Evaluation of different methods

Exosomes are small RNA containing vesicles of endocytic origin, which can take part in cell-to-cell communication partly by the transfer of exosomal RNA between cells. Exosomes are released by many cells and can also be found in several biological fluids including blood plasma and breast milk. Exosomes differ compared to their donor cells not only in size but also in RNA, protein and lipid composition. The aim of the current study was to determine the optimal RNA extraction method for analysis of exosomal RNA, to support future studies determining the biological roles of the exosomal RNA. [2]

RNA isolation from recalcitrant plant tissue

The isolation of high-quality RNA from various tissues (leaves, pedicels, glandular trichomes) of garden geranium (Pelargonium xhortorum) using various published methods is difficult due to numerous oxidizing compounds. A new RNA extraction method was developed through the combination and modification of two separate procedures (Rochester et al., 1986; Manning 1991). In addition to geranium tissues, this method is successful when used with other recalcitrant tissues such as mature needles of white pine (Pinus strobus) and mature leaves of poinsettia (Euphorbia pulcherrima). RNA quality was judged by spectrophotometric readings, denaturing agarose gels, and successful reverse transcription. [3]

Isolation, Cloning and Co-expression of Hepatitis C Virus Envelope Proteins: As Potential HCV Detecting Antigens

Aim: Approximately 3% of the world population is infected with Hepatitis C virus (HCV) which is the main cause of chronic liver disease. Blood transfusion is thought to be the leading cause of global epidemic of HCV. The envelope proteins E1 and E2 are involved in the early stages of the virus life cycle. These proteins have a major role in binding to receptors on the cell surface, fusion and integration of the virus into the host cell. Considering the potency of E1 and E2 in the development of diagnostic methods, the aim of our present study was co-expression of recombinant envelope proteins in eukaryotic HEK293 (human embryonic kidney) cells.

Methods: The viral genomic RNA was used for cDNA (complementary DNA) synthesis. Isolation of HCV envelope proteins coding fragment was performed using cDNA and specific primers. The target gene was cloned into pcDNA3.1 expression vector, and transfected into HEK293 cells, an expression host. Accuracy of the cloning and expression was confirmed using PCR and Western blot analysis.

Results: The isolation and cloning of the gene fragment encoding the E1 and E2 proteins was successful. Co-expression of these proteins was confirmed using monoclonal antibodies specific for each protein.

Conclusion: This study showed that HEK293 host cell is suitable for the expression of hepatitis C virus E1 and E2 coding gene. These proteins can be used in numerous virological studies and detection of HCV infection. [4]

Isolation and Molecular Characterization of Lactic Acid Bacteria Isolated from Fresh Fruits and Vegetables Using Nested PCR Analysis

Aims: The study investigated the diversity and identities of Lactic Acid Bacteria (LAB) isolated from different fresh fruits and vegetables using Molecular Nested PCR analysis with the view of identifying LAB with anti-microbial potentials.

Study Design: Nested PCR approach was used in this study employing universal 16S rRNA gene primers in the first round PCR and LAB specific Primers in the second round PCR with the view of generating specific Nested PCR products for the LAB diversity present in the samples.

Place and Duration of Study: Biotechnology Centre of Federal University of Agriculture, Abeokuta, Ogun State, Nigeria, between January 2011 and February 2012.

Methodology: Forty Gram positive, catalase negative strains of LAB were isolated from fresh fruits and vegetables on Man Rogosa and Sharpe agar (Lab M) using streaking method. Standard molecular methods were used for DNA extraction (Norgen Biotek kit method, Canada), Polymerase Chain Reaction (PCR) Amplification, Electrophoresis, Purification and Sequencing of generated Nested PCR products (Macrogen Inc., USA).

Results: The partial sequences obtained were deposited in the database of National Centre for Biotechnology Information (NCBI). Isolates were identified based upon the sequences as Weissella cibaria (5 isolates, 27.78%), Weissella kimchi (5, 27.78%), Weissella paramensenteroides (3, 16.67%), Lactobacillus plantarum (2, 11.11%), Pediococcus pentosaceus (2, 11.11%) and Lactobacillus pentosus (1, 5.56%) from fresh vegetable; while Weissella cibaria (4, 18.18%), Weissella confusa (3, 13.64%), Leuconostoc paramensenteroides (1, 4.55%), Lactobacillus plantarum (8, 36.36%), Lactobacillus paraplantarum (1, 4.55%) and Lactobacillus pentosus (1, 4.55%) were identified from fresh fruits.

Conclusion: This study shows that potentially LAB can be quickly and holistically characterized by molecular methods to specie level by nested PCR analysis of the bacteria isolate genomic DNA using universal 16S rRNA primers and LAB specific primer. [5]

Reference

[1] Chomczynski, P. and Sacchi, N., 2006. The single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction: twenty-something years on. Nature protocols, 1(2), pp.581-585.

[2] Eldh, M., Lötvall, J., Malmhäll, C. and Ekström, K., 2012. Importance of RNA isolation methods for analysis of exosomal RNA: evaluation of different methods. Molecular immunology, 50(4), pp.278-286.

[3] Schultz, D.J., Craig, R., Cox-Foster, D.L., Mumma, R.O. and Medford, J.I., 1994. RNA isolation from recalcitrant plant tissue. Plant Molecular Biology Reporter, 12(4), pp.310-316.

[4] Mohammadipour, M., Ahangari, G. and Sadeghizadeh, M. (2015) “Isolation, Cloning and Co-expression of Hepatitis C Virus Envelope Proteins: As Potential HCV Detecting Antigens”, Journal of Advances in Medicine and Medical Research, 9(11), pp. 1-8. doi: 10.9734/BJMMR/2015/16720.

[5] Emerenini, E. C., Afolabi, O. R., Okolie, P. I. and Akintokun, A. K. (2013) “Isolation and Molecular Characterization of Lactic Acid Bacteria Isolated from Fresh Fruits and Vegetables Using Nested PCR Analysis”, Microbiology Research Journal International, 3(3), pp. 368-377. doi: 10.9734/BMRJ/2013/2520.

Latest News on Metalloprotease: Dec 2020

Induction of Cell Migration by Matrix Metalloprotease-2 Cleavage of Laminin-5

Structural changes in the extracellular matrix are necessary for cell migration during tissue remodeling and tumor invasion. Specific cleavage of laminin-5 (Ln-5) by matrix metalloprotease–2 (MMP2) was shown to induce migration of breast epithelial cells. MMP2 cleaved the Ln-5 γ2 subunit at residue 587, exposing a putative cryptic promigratory site on Ln-5 that triggers cell motility. This altered form of Ln-5 is found in tumors and in tissues undergoing remodeling, but not in quiescent tissues. Cleavage of Ln-5 by MMP2 and the resulting activation of the Ln-5 cryptic site may provide new targets for modulation of tumor cell invasion and tissue remodeling. [1]

Nitric Oxide Activates Metalloprotease Enzymes in Articular Cartilage

Nitric oxide (NO.) is a multifunctional messenger molecule generated by a family of enzymes, collectively termed the nitric oxide synthases. We investigated the role of NO. in the modulation of two metal-dependent proteolytic enzymes (collagenase and stromelysin) which are activated during inflammatory and infective arthritis. The inflammatory mediators interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and the bacterial cell wall fragment endotoxin, induced both nitric oxide synthase activity and stromelysin and collagenase activity in whole cell preparations and in conditioned media from explants of bovine and human cartilage. Both NO2- (the stable end-product of NO.) and metalloprotease activity were inhibited by competitive inhibitors of nitric oxide synthase. The NO. donor, S-nitroso-N-acetyl-D,L-penicillamine (SNAP) also induced metalloprotease activity in a dose-dependent fashion. These data provide evidence that NO. plays a regulatory role in the activation of metal-dependent proteases in articular chondrocytes and cartilage. [2]

A Novel Proteolytic Cleavage Involved in Notch Signaling: The Role of the Disintegrin-Metalloprotease TACE

The Notch1 receptor is presented at the cell membrane as a heterodimer after constitutive processing by a furin-like convertase. Ligand binding induces the proteolytic release of Notch intracellular domain by a γ-secretase-like activity. This domain translocates to the nucleus and interacts with the DNA-binding protein CSL, resulting in transcriptional activation of target genes. Here we show that an additional processing event occurs in the extracellular part of the receptor, preceding cleavage by the γ-secretase-like activity. Purification of the activity accounting for this cleavage in vitro shows that it is due to TACE (TNFα-converting enzyme), a member of the ADAM (a disintegrin and metalloprotease domain) family of metalloproteases. Furthermore, experiments carried out on TACE−/− bone marrow–derived monocytic precursor cells suggest that this metalloprotease plays a prominent role in the activation of the Notch pathway. [3]

Characterization of Trypsin-Like Serine Protease from Lethocerus indicus Salivary Venom and its Cytotoxic Effect against Human Epidermoid Carcinoma Cell, A431

Objective: Lethocerus indicus salivary venom characterization and evaluation of extracellular degradation activity and cytotoxic effect against native human collagen type 1 and epidermoid carcinoma cell, A431.

Method: Salivary venom extract was collected from adult insects by injecting 2% pilocarpine of 50 µml. Enzyme presence was detected by the apiZYM assay. The proteolytic activity was tested by the photometric and zymogram methods using specific fluorescent substrates and inhibitors. The cytotoxic activity was determined by the MTT assay and Trypan blue exclusion method. Apoptosis induction was observed using AO/EB staining solution. Digestion of extracellular matrix protein was detected against native human type I collagen.

Result: L. indicus salivary venom presents amylases, proteases, carbohydrases, phosphatases and lipases. Among them, protease enzyme showed highest composition. The highest rate of proteolytic activity observed at pH 8 in 35ºC (100 %). Serine proteases present predominantly in salivary venom. Cysteine and metalloproteases are also detected. The activation energy of salivary venom is 49.86 kJ. Use of serine inhibitor, PMSF inhibited 92.77% which indicated that the maximum activity was due to serine protease. Detection of trypsin-like protease was confirmed by using PMSF and TLCK with specific substrate, BApNA. It shows significant inhibitions, 82% and 78% respectively suggesting maximum influence in salivary venom. Degradation of the fibrillar native state collagen Type I into 8 smaller peptide bands showed it importance in medical application. IC50 concentration of venom that induces cytotoxicity in epidermoid carcinoma cells, A431was 2.3 µg/ml only. It gives prominent apoptotic features such as cytoplasmic membrane blebbing, nuclear contraction, nuclear fragmentation and contact inhibition.

Conclusion: We suggest that further investigation of the venom will lead to identification of active compound in L. indicus salivary venom for its potential use in therapeutic application. [4]

The Distribution of Matrix Metalloproteinase-2, Tissue Inhibitor of Metalloproteinase-2 and Tissue Inhibitor of Metalloproteinase-4 in Psoriatic Skin

Aims: To evaluate the appearance and distribution of matrix metalloproteinase-2 (MMP-2), tissue inhibitor of metalloproteinase-2 (TIMP-2) and tissue inhibitor of metalloproteinase-4 (TIMP-4) in lesional skin biopsies of psoriasis patients.

Study Design: Observational study.

Place and Duration of Study: Institute of Anatomy and Anthropology and Department of Infectology and Dermatology, RÄ«ga Stradiņš University, between September 2013 and June 2014.

Methodology: We included 40 patients (31 men, 9 women; age range 18-70 years) with Psoriasis vulgaris, with present characteristic psoriatic eruptions in typical localization sites and no treatment received. Skin samples were obtained using routine punch biopsy method. 10 clinically healthy skin samples obtained during nevus excision procedure were used as control material. All tissue specimens were stained with hematoxylin and eosin and by immunohistochemistry for MMP-2, TIMP-2 and TIMP-4. The intensity of staining was graded semiquantitatively. Spearman’s rank correlation coefficient was calculated.

Results: In psoriasis patients numerous MMP-2-containing keratinocytes were found in epidermis, MMP-2 positive dermal fibroblasts and inflammatory cells varied from few to abundant. Few epidermal cells and moderate to numerous dermal cells contained TIMP-2. Moderate to numerous epidermal and dermal cells contained TIMP-4. Statistically significant strong positive correlation was found between MMP-2 in epidermis and dermis (Spearman’s rank correlation coefficient = .878, P = .000). Statistically significant moderate positive correlation was found between TIMP-2 and TIMP-4 in dermis (Spearman’s rank correlation coefficient = .639, P = .000) and between TIMP-2 in epidermis and dermis (Spearman’s rank correlation coefficient = .564, P = .000).

Conclusion: TIMP-4 seems to be most important inhibitor of psoriatic skin degeneration, richly raised by MMP-2. Its moderate correlation with TIMP-2 proves involvement of other tissue inhibitors in the degeneration inhibition and gives evidence about possible patterning between the tissue inhibitors of metalloproteinases. [5]

Reference

[1] Giannelli, G., Falk-Marzillier, J., Schiraldi, O., Stetler-Stevenson, W.G. and Quaranta, V., 1997. Induction of cell migration by matrix metalloprotease-2 cleavage of laminin-5. Science, 277(5323), pp.225-228.

[2] Murrell, G.A., Jang, D. and Williams, R.J., 1995. Nitric oxide activates metalloprotease enzymes in articular cartilage. Biochemical and biophysical research communications, 206(1), pp.15-21.

[3] Brou, C., Logeat, F., Gupta, N., Bessia, C., LeBail, O., Doedens, J.R., Cumano, A., Roux, P., Black, R.A. and Israël, A., 2000. A novel proteolytic cleavage involved in Notch signaling: the role of the disintegrin-metalloprotease TACE. Molecular cell, 5(2), pp.207-216.

[4] Debaraj, H., Shantibala, T., K. Lokeshwari, R. and Giri, S. (2014) “Characterization of Trypsin-Like Serine Protease from Lethocerus indicus Salivary Venom and its Cytotoxic Effect against Human Epidermoid Carcinoma Cell, A431”, Biotechnology Journal International, 4(9), pp. 990-1010. doi: 10.9734/BBJ/2014/12217.

[5] Sidhom, E., Pilmane, M. and Kisis, J. (2015) “The Distribution of Matrix Metalloproteinase-2, Tissue Inhibitor of Metalloproteinase-2 and Tissue Inhibitor of Metalloproteinase-4 in Psoriatic Skin”, Journal of Advances in Medicine and Medical Research, 8(10), pp. 883-890. doi: 10.9734/BJMMR/2015/18206.

Latest News on Induced Oxidative Stress: Dec 2020

Advances in metal-induced oxidative stress and human disease

Detailed studies in the past two decades have shown that redox active metals like iron (Fe), copper (Cu), chromium (Cr), cobalt (Co) and other metals undergo redox cycling reactions and possess the ability to produce reactive radicals such as superoxide anion radical and nitric oxide in biological systems. Disruption of metal ion homeostasis may lead to oxidative stress, a state where increased formation of reactive oxygen species (ROS) overwhelms body antioxidant protection and subsequently induces DNA damage, lipid peroxidation, protein modification and other effects, all symptomatic for numerous diseases, involving cancer, cardiovascular disease, diabetes, atherosclerosis, neurological disorders (Alzheimer’s disease, Parkinson’s disease), chronic inflammation and others. The underlying mechanism of action for all these metals involves formation of the superoxide radical, hydroxyl radical (mainly via Fenton reaction) and other ROS, finally producing mutagenic and carcinogenic malondialdehyde (MDA), 4-hydroxynonenal (HNE) and other exocyclic DNA adducts. On the other hand, the redox inactive metals, such as cadmium (Cd), arsenic (As) and lead (Pb) show their toxic effects via bonding to sulphydryl groups of proteins and depletion of glutathione. Interestingly, for arsenic an alternative mechanism of action based on the formation of hydrogen peroxide under physiological conditions has been proposed. A special position among metals is occupied by the redox inert metal zinc (Zn). Zn is an essential component of numerous proteins involved in the defense against oxidative stress. It has been shown, that depletion of Zn may enhance DNA damage via impairments of DNA repair mechanisms. In addition, Zn has an impact on the immune system and possesses neuroprotective properties. The mechanism of metal-induced formation of free radicals is tightly influenced by the action of cellular antioxidants. Many low-molecular weight antioxidants (ascorbic acid (vitamin C), alpha-tocopherol (vitamin E), glutathione (GSH), carotenoids, flavonoids, and other antioxidants) are capable of chelating metal ions reducing thus their catalytic acitivity to form ROS. A novel therapeutic approach to supress oxidative stress is based on the development of dual function antioxidants comprising not only chelating, but also scavenging components. Parodoxically, two major antioxidant enzymes, superoxide dismutase (SOD) and catalase contain as an integral part of their active sites metal ions to battle against toxic effects of metal-induced free radicals. The aim of this review is to provide an overview of redox and non-redox metal-induced formation of free radicals and the role of oxidative stress in toxic action of metals. [1]

Environmentally induced oxidative stress in aquatic animals

Reactive oxygen species (ROS) are an unenviable part of aerobic life. Their steady-state concentration is a balance between production and elimination providing certain steady-state ROS level. The dynamic equilibrium can be disturbed leading to enhanced ROS level and damage to cellular constituents which is called “oxidative stress”. This review describes the general processes responsible for ROS generation in aquatic animals and critically analyses used markers for identification of oxidative stress. Changes in temperature, oxygen levels and salinity can cause the stress in natural and artificial conditions via induction of disbalance between ROS production and elimination. Human borne pollutants can also enhance ROS level in hydrobionts. The role of transition metal ions, such as copper, chromium, mercury and arsenic, and pesticides, namely insecticides, herbicides, and fungicides along with oil products in induction of oxidative stress is highlighted. Last years the research in biology of free radicals was refocused from only descriptive works to molecular mechanisms with particular interest to ones enhancing tolerance. The function of some transcription regulators (Keap1–Nrf2 and HIF-1α) in coordination of organisms’ response to oxidative stress is discussed. The future directions in the field are related with more accurate description of oxidative stress, the identification of its general characteristics and mechanisms responsible for adaptation to the stress have been also discussed. The last part marks some perspectives in the study of oxidative stress in hydrobionts, which, in addition to classic use, became more and more popular to address general biological questions such as development, aging and pathologies. [2]

Exercise-induced oxidative stress.

The role of exercise in free radical processes is not clear; however, recent evidence suggests that elevated oxygen consumption may increase free radical activity. Direct measurement of free radical signals can be made by electron spin resonance and indirect measures include mitochondrial membrane damage, conjugated dienes, hydroperoxides, thiobarbituric acid reactive substances, short chain hydrocarbons, and oxidized nucleosides. Although exact levels are not known, the type, duration, and intensity of exercise affect biomarkers of free radical activity, as does one’s training status. Oxidative stress associated with exercise-induced free radical activity seems to be better tolerated by trained subjects exercising at moderate intensity. [3]

Protective Potential of Grape Seed Proanthocyandins Extract against Glivec (Imatinib Mesylate) Induced Liver Toxicity and Oxidative Stress in Male Rats

Objectives: Glivec (Imatinib mesylate) an antineoplastic chemotherapeutic agent used in the treatment of many types of cancer. The current study examines the hepatoprotective potential of grape seed proanthocyandins extract (GSPE) against Glivec induced oxidative stress and toxicity in male albino rats.

Materials and Methods: A total of 40 male albino rats were equally divided into four groups; group 1 was control, group 2 was GSPE group (rats received orally GSPE by stomach tube {50 mg/kg BW/twice a week} for four week), group 3 was Glivec group (rats were injected intraperitoneally with Glivec {1 mg /kg B W/twice a week} for four weeks) and group 4 was rats treated with GSPE plus Glivec for four weeks.

Results: LD50 was calculated for Glivec in rats (estimated at 598 mg/kg), presenting confidence limits between 588 and 612 mg/kg body weight. A significant increase in the liver TBARS and aspartate aminotransferase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), and γ-glutamyltransferase (GGT) activities in Glivec group when compared with the control group. On the other hand; a significant decrease in the serum albumin, globulin, total protein, liver superoxide dismutase activity (SOD), catalase (CAT), glutathione S-trasferase (GST) and reduced glutathione (GSH) levels in Glivec group when compared with the control group. Administration of GSPE with Glivec caused a protective and ameliorative effect against Glivec induced liver toxicity.

Conclusions: Treatment with GSPE has a promising role for ameliorating the oxidative stress and hepatic injury induced by Glivec. [4]

Neuroprotective Effect of Convolvulus pluricaulis Methanol Extract on Hydrogen Peroxide Induced Oxidative Stress in Human IMR32 Neuroblastoma Cell Line

Aims: The present study aimed to evaluate and ascertain the protective role of methanolic/ethanolic/water extracts of Convolvulus pluricaulis against H2O2 induced cytotoxicity in IMR32 Neuroblastoma cell line as model system and identify the factor responsible for the protective effect.

Study Design:  Experimental study.

Place and Duration of Study: Department of Molecular Biology and Biochemistry, Guru Nanak Dev University, Amritsar & Department of Biotechnology, DAV College, Amritsar, PuCPab, between August 2010 and March 2012.

Methodology: Firstly, cytotoxic dose of H2O2 ­and non-toxic dose of methanolic, ethanolic and water extracts of C. pluricaulis (CP-MEx, CP-EEx and CP-WEx respectively) was determined by MTT assay. Protective effect of CP-MEx, CP-EEx and CP-WEx was determined using quercetin as a positive control. The expression of IMR32 cytoskeletal marker, Neurofilament (NF-200) and stress markers, Heat shock protein (HSP70) and (glucose regulated protein 75, Grp75) Mortalin studied by immunofluorescence and RT-PCR results. The level of antioxidant enzymes catalase, superoxide dismutase, glutathione peroxidase, direct scavenger of free radicals, Glutathione and lipid peroxidation were analysed by their standard procedures.

Results: The results showed that quercetin, CP-MEx, CP-EEx and CP-WEx displayed cytoprotective activity in IMR32 cells. Out of tested extracts CP-MEx significantly decreased hydrogen peroxide-induced cell death. Significant decrease in NF-200, HSP70 and Mortalin expression was observed in CP-MEx+H2O2 treated cultures as compared to H2O2 treated. Catalase, superoxide dismutase, glutathione peroxidase, Glutathione levels significantly increased in Quercetin and CP-MEx treated cultures. Lipid peroxidation was significantly decreased in both Quercetin and CP-MEx treated cultures.

Conclusions: The present work establishes the protective effect of CP-MEx on IMR 32 Human Neuroblastoma cell line which is as much as by quercetin. The cytoprotective effect of CP-MEx was due to induction of antioxidant machinery of the cell hence holds therapeutic value in the treatment and/or prevention of neurodegenerative disorders of oxidative stress. [5]

Reference

[1] Jomova, K. and Valko, M., 2011. Advances in metal-induced oxidative stress and human disease. Toxicology, 283(2-3), pp.65-87.

[2] Lushchak, V.I., 2011. Environmentally induced oxidative stress in aquatic animals. Aquatic toxicology, 101(1), pp.13-30.

[3] Alessio, H.M., 1993. Exercise-induced oxidative stress. Medicine and science in sports and exercise, 25(2), pp.218-224.

[4] Al-Rasheed, N. M., El-Masry, T. A., Tousson, E., Hassan, H. M. and Al-Ghadeer, A. (2017) “Protective Potential of Grape Seed Proanthocyandins Extract against Glivec (Imatinib Mesylate) Induced Liver Toxicity and Oxidative Stress in Male Rats”, Annual Research & Review in Biology, 20(6), pp. 1-9. doi: 10.9734/ARRB/2017/37766.

[5] Dhuna, K., Dhuna, V., Bhatia, G., Singh, J. and Singh Kamboj, S. (2012) “Neuroprotective Effect of Convolvulus pluricaulis Methanol Extract on Hydrogen Peroxide Induced Oxidative Stress in Human IMR32 Neuroblastoma Cell Line”, Biotechnology Journal International, 2(4), pp. 192-210. doi: 10.9734/BBJ/2012/1655.

Latest News on Enzymes : Dec 2020

Soil Enzymes

Nutrient cycling in soils involves biochemical, chemical, and physiochemical reactions, with the biochemical processes being mediated by microorganisms, plant roots, and soil animals. Enzymes are denatured by elevated temperature and extreme pH. The two most remarkable properties of enzymes are their specificity and catalytic efficiency, and it is in these properties that enzymes differ most strikingly from simple catalysts. Temperature affects enzyme activity in various ways; the number of factors controlling the effect of pH on an enzyme‐catalyzed reaction is even greater. The subject of enzyme inhibition requires special consideration, because soils receive a variety of organic and inorganic chemicals. Soil is a living system where all biochemical activities proceed through enzymatic processes. The solutions of the substrates used for assay of phosphomonoesterases and phosphodiesterase are stable for several days if stored in a refrigerator. The activities of several other enzymes have been detected in soils and methods have been developed for their assays. [1]

Lipolytic Enzymes

A major group of lipolytic enzymes typified by pancreatic lipase consists of nonspecific esterases probably of the serinehistidine type. Another group, the phospholipases 2 of exocrine glands, are calcium activated and have exacting stereo-specific substrate requirements. The enzymes of both groups hydrolyze water-insoluble esters. They must not only adsorb to oil—water or micelle—water interfaces but also must position their active sites toward the matrix (oil droplet, micelle, or membrane) in which the substrate molecules are imbedded, the “supersubstrate” It is postulated that lipolytic enzymes are hydrolases that have developed supersubstrate binding sites for attachment and orientation toward lipids. Such a binding site, which is topographically distinct from the substrate binding sites of the reactive center, may have hydrophobic or electrostatic character. [2]

Antioxidant enzymes and human diseases

Objectives: To describe the importance of the antioxidant enzymes superoxide dismutase, glutathione peroxidase, and catalase working together in human cells against toxic reactive oxygen species, their relationship with several pathophysiologic processes and their possible therapeutic implications.

Conclusions: Reactive oxygen species (ROS) are involved in the cell growth, differentiation, progression, and death. Low concentrations of ROS may be beneficial or even indispensable in processes such as intracellular signaling and defense against micro-organisms. Nevertheless, higher amounts of ROS play a role in the aging process as well as in a number of human disease states, including cancer, ischemia, and failures in immunity and endocrine functions. As a safeguard against the accumulation of ROS, several non-enzymatic and enzymatic antioxidant activities exist. Therefore, when oxidative stress arises as a consequence of a pathologic event, a defense system promotes the regulation and expression of these enzymes. [3]

Determination of SOD, POD, PPO and CAT Enzyme Activities in Rumex obtusifolius L.

Aims: The purpose of this study was to measure antioxidant enzyme (polyphenol oxidase, peroxidase, catalase and superoxide dismutase) activities of crude extract of Rumex obtusifolius L. in order to gain insight about this plant’s antioxidant potential.

Study Design: The study was composed of the collection of plant material, extractions of the antioxidant enzymes, activity measurements of the enzymes and finally evaluation of the experimental results.

Place and Duration of Study: Department of Chemistry (biochemistry laboratories), Faculty of Science and Arts of Sakarya University, between June 2015 and July 2015.

Methodology: Enzymatic antioxidant activity of this plant was investigated by carrying out catalase, superoxide dismutase, peroxidase and polyphenol oxidase enzyme activity assays. Enzyme activities of the crude extract were measured by using spectrophotometric method. Optimum pH and temperature values of each enzyme were also determined for measurement of enzyme activities in ideal conditions.

Results: Finally, our results showed that Rumex obtusifolius L. crude extract had good activity for all the enzymatic procedures tested. The activity levels of enzymatic antioxidants polyphenol oxidase, peroxidase, catalase and superoxide dismutase of the plant were found to be 12.8; 195.2; 38.7; 11.6 EU/mL, respectively. Optimum pH and temperature values of all the enzymes (except PPO: optimum temperature 30°C) tested were also found to be 7.0 and 25°C, respectively.

Conclusion: Our results demonstrate that this edible plant, Rumex obtusifolius L., might be a potential source of natural antioxidants with good antioxidant enzyme capacity. [4]

Influence of Soil Ameliorants, Manures and Fertilizers on Bacterial Populations, Enzyme Activities, N Fixation and P Solubilization in Peanut Rhizosphere under Lateritic Soil

The aim of this study was to investigate relative efficacy of different organic wastes like farmyard manure (FYM) and water hyacinth (WH) and industrial wastes like paper factory sludge (PFS) on balancing with chemical fertilizers (CF) along with soil ameliorants viz., lime (L) or rice husk ash (RHA), another industrial waste, on dry matter production and biological properties of the rhizosphere soil of peanut (Arachis hypogaea), grown as intercrop with sabai grass (Eulaliopsis binata) in acid lateritic soil. Population of symbiotic nitrogen fixing and phosphorus solubilizing bacteria, activity of dehydrogenase and phosphatase enzymes (i.e. acid and alkaline phosphomonoesterases), nitrogen accumulation in nodules and phosphorus solubilizing power of rhizosphere soil were measured after 25, 50, 75 and 100 days after sowing (DAS) of peanut for two years. Results showed significant effects of nutrient sources and growth stages of the crop on the microbial activities. Higher values of all the biological properties and plant growth parameters were recorded significantly under the integrated application of CF and any of the organic or industrial wastes over sole application of CF. Among three organic or industrial wastes WH was superior to others regarding microbial activities at 25 DAS, whereas PFS became superior at 50, 75 and 100 DAS. Application of lime or RHA improved the activity of dehydrogenase and alkaline phosphomonoesterase enzymes, while decreased acid phosphomonoesterase activity. This study revealed that integrated application of organic or industrial wastes, soil ameliorants and inorganic fertilizer, could improve the biological properties of an acid lateritic soil as well as the dry matter production of peanut, intercropped with sabai grass under lateritic soil. [5]

Reference

[1] Tabatabai, M.A., 1994. Soil enzymes. Methods of Soil Analysis: Part 2 Microbiological and Biochemical Properties, 5, pp.775-833.

[2] Brockerhoff, H., 1974. Lipolytic enzymes.

[3] Matés, J.M., Pérez-Gómez, C. and De Castro, I.N., 1999. Antioxidant enzymes and human diseases. Clinical biochemistry, 32(8), pp.595-603.

[4] Alici, E. and Arabaci, G. (2016) “Determination of SOD, POD, PPO and CAT Enzyme Activities in Rumex obtusifolius L.”, Annual Research & Review in Biology, 11(3), pp. 1-7. doi: 10.9734/ARRB/2016/29809.

[5] Basu, M., Bhadoria, P. B. S. and Mahapatra, S. C. (2011) “Influence of Soil Ameliorants, Manures and Fertilizers on Bacterial Populations, Enzyme Activities, N Fixation and P Solubilization in Peanut Rhizosphere under Lateritic Soil”, Microbiology Research Journal International, 1(1), pp. 11-25. Available at: https://www.journalmrji.com/index.php/MRJI/article/view/5983 (Accessed: 30November2020).

Latest News on Nutrient Management : Nov 2020

Biogeochemical nutrient cycles and nutrient management strategies

Nutrient loading by riverine input into estuarine systems has increased by 6–50 times for the N load from pristine conditions to present, whereas a 18–180 times increase has been observed in the P load. Reductions in the ratio of N to P delivery has also occurred with time. In a review of nutrient limitation in estuarine systems, it is shown that many estuarine systems display P limitation in the spring, switching to N limitation in the summer with some estuaries displaying dissolved silicate limitation of the spring diatom bloom. Historical and recent changes in nutrient loading and their effect on nutrient limitation have intensified the debate on the control of nutrient delivery to estuaries from both agricultural and point sources, and as to what nutrient (N or P) should be managed for in estuarine systems. It is hypothesized that potential reductions in P may help oxygen depletion especially in deep estuaries and reduce fast growing macrophytes such as Ulva sp., although P reductions probably will have little effect on summer chlorophyll concentrations, an important recreational management goal. Reductions in N loading should reduce summer chlorophyll concentrations and improve the conditions for submerged aquatic vegetation and thus improve ecosystem functioning. Finally, if only P reductions are pursued, that is if we are able to reduce P such that it is limiting year around in estuarine systems, it is likely that the export of N from estuarine systems would increase to the bordering N-limited marine systems, thus only exporting the problem of enhanced production with eutrophication. [1]

Approaches and uncertainties in nutrient budgets: implications for nutrient management and environmental policies

Nutrient budgets of agroecosystems are constructed either (i) to increase the understanding of nutrient cycling, (ii) as performance indicator and awareness raiser in nutrient management and environmental policy, or (iii) as regulating policy instrument to enforce a certain nutrient management policy in practice. This paper explores nutrient budgeting approaches and summarizes sources of uncertainty associated with these approaches. Possible implications of uncertainties associated with the different methodologies and approaches for nutrient management and environmental policy are discussed. Three types of nutrient budgets have been distinguished, i.e. farm-gate, soil surface and soil systems budgets. A farm-gate budget is the most integrative measure of environmental pressure, and seems most suitable as environmental performance indicator. A soil surface budget is appropriate for estimating the net loading of the soil with nutrients. Soil system budgets account for nutrient inputs and outputs, recycling of nutrients within the system, nutrient loss pathways and changes in soil nutrient pools; it is the most detailed budget and provides detailed information for nutrient management. Case studies for the dairy farm De Marke indicate that the three budgeting approaches supplement each other. The accuracy and precision of the nutrient budget depend on budgeting approach, data acquisition strategy and type of agroecosystem. There is often a considerable amount of uncertainty in the nutrient budget, due to various possible biases and errors, notably in the partitioning of nutrient losses. Possible sources of biases are personal bias, sampling bias, measurement bias, data manipulation bias and fraud. Sources of errors are sampling and measurement errors. Both biases and errors in nutrient budget estimates may lead to confusion and wrong conclusions. Yet, there is little published evidence that uncertainties are taken into account in decision making. Uncertainties are usually smaller for a farm-gate budget than for a soil surface budget. Therefore, farm-gate budgets are preferred over soil surface budgets as policy instrument. Quantifying uncertainties requires: (a) system identification and analysis, (b) classification of uncertainties, (c) specification of distributions of probabilities of the various sources using Monte Carlo simulation, and (d) monitoring of the nutrient pools, inputs and outputs over time. Analyses of uncertainties in nutrient budgets may provide information about the weakest chain in establishing agri-environmental cause–effect relationships and, therefore, may assist to better focus research efforts. Data for the nitrogen budget of The Netherlands indicate relatively large uncertainties for the items denitrification and leaching, with coefficients of variation >30%. In conclusion, there is a need for standard procedures and guidelines in nutrient budgeting and uncertainty analyses, to improve the confidence in and applicability of nutrient budgets. [2]

Development of phosphorus indices for nutrient management planning strategies in the United States

Phosphorus (P), an essential nutrient for crop and livestock production, can accelerate freshwater eutrophication, now one of the leading water quality impairments in the United States. In response, the U.S. Department of Agriculture and the Environmental Protection Agency proposed a new nutrient management policy, now addressing P as well as nitrogen (N), which each state must enact by 2008. There are three approaches that address P—agronomic soil test P recommendations, environmental soil test P thresholds, and a P index to rank fields according to their vulnerability to potential P loss. There are many versions of the P index now in use, demonstrating the robustness and flexibility of the indexing framework to better target remedial measures. Of the three P-based approaches, the P indexing approach has been most widely adopted with 47 states using this approach to target P management. This paper charts the development of the indexing approach, which ranks site vulnerability to P loss by accounting for source (soil test P, fertilizer, and manure management) and transport factors (erosion, runoff, leaching, and connectivity to a stream channel) and outlines modifications made among states that reflect local conditions and policy. Additional factors include flooding frequency, STP modifiers (texture, pH, P sorption, reactive aluminum [Al]), conservation practices, and priority of receiving waters. While computation of the final index value is additive in 20 states, 17 multiply source and transport factors to define critical source areas. Most states (47) have maintained the original indexing approach of assessing site vulnerability to P loss, with indices in three states quantifying P loss. We demonstrate using three management scenarios (changing the time of applied manure, riparian buffer establishment, and reduced feed P ration) that overall P index ratings can be decreased, giving farmers more options for manure management than by simply reducing application rates. [3]

Integrated Nutrient Management; Option for Improvement on Soil Chemical Properties, Growth and Yield of Cocoyam (Colocasia esculenta)

The response of cocoyam (Colocasia esuclenta (L.) Schott) and soil chemical properties to different manure sources were studied under field conditions in 2014 and 2015 cropping season at Federal College of Agriculture, Ishiagu, Ebonyi State, a derived savanna zone of Southeastern Nigeria. Randomized complete block design (RCBD) was used to study these soil amendments. Ten(10) treatments; 10t/ha poultry dropping (PD), 200 kg/ha NPK 15:15:15 fertilizer (NPK), 5 t/ha rice husk ash (RHA), 10 t/ha rice husk dust (RHD), 5t/ha PD + 2.5 t/ha RHA, 5 t/ha PD + 5 t/ha RHD, 100 kg/ha NPK + 2.5 t/ha RHA, 100 kg/ha NPK + 5t/ha RHD, 66.67 kg/ha NPK + 3.33 t/ha RHD + 1.67 t/ha RHA and control, were used in the study which were replicated three times. Soil chemical properties evaluated were pH, organic carbon, exchangeable bases (K+, Ca2+, Mg 2+ and Na+) and exchangeable acidity. Others include CEC, total nitrogen, available phosphorus and base saturation while cocoyam yield was measured at harvest. The results showed that soil pH, soil organic carbon (SOC), cation exchange capacity (CEC), exchangeable acidity (EA), BS and available phosphorous were significantly affected by the application of the amendments, while total nitrogen shows non-significant effect. Cocoyam cormels and corms yield were significantly (p < 0.05) increased in all the soil amended plots than the control. Generally, the results implied that the integration of organic and inorganic amendments stand better chance of improving soil fertility indices and the crop yield than their sole forms in the study. [4]

Response of Potato Crop to Integrated Nutrient Management in the Indo-Gangetic Alluvial Soils of West Bengal, India

A field experiment was carried out to investigate the influence of Integrated Nutrient Management (INM) on nutrient uptake and yield of potato (Solanum tuberosum L.) in the Gangetic alluvium belt of West Bengal during two consecutive years i.e. 2011-12 to 2012-13. The experiment was laid out in a randomized complete block design with three replications. Different levels and combinations of organic (crop residue, farm yard manure and bio-fertilizer) and chemical fertilizers were tested. Plots receiving only organic manures reported least amount of nutrient uptake (36.00-44.65, 7.57-11.0 and 44.66-72.66 kg·ha-1 for N, P and K respectively). The nutrient uptake was significantly higher when chemical fertilizers were applied. Maximum response was found (up-to 161, 221 and 354% increment for N, P and K respectively compared to control) when different sources were combined together. Strong correlation has been found between nutrient uptake, tuber dry weight, specific gravity, ascorbic acid and reducing sugar content suggesting role of nutrients uptake on tuber yield and quality. With increasing nutrient uptake, up to 80% increment in yield was found from those plots. It was noted from our experiment that sole use of organics was not effective enough for supplying nutrients. However, the integration of organic fertilizers with chemical ones had significantly improved soil and crop health, providing better crop yield and quality. [5]

Reference

[1] Conley, D.J., 1999. Biogeochemical nutrient cycles and nutrient management strategies. In Man and River Systems (pp. 87-96). Springer, Dordrecht.

[2] Oenema, O., Kros, H. and de Vries, W., 2003. Approaches and uncertainties in nutrient budgets: implications for nutrient management and environmental policies. European Journal of Agronomy, 20(1-2), pp.3-16.

[3] Sharpley, A.N., Weld, J.L., Beegle, D.B., Kleinman, P.J., Gburek, W.J., Moore, P.A. and Mullins, G., 2003. Development of phosphorus indices for nutrient management planning strategies in the United States. Journal of Soil and Water Conservation, 58(3), pp.137-152.

[4] Nwite, J. C., Keke, C. I. and Okereke, P. O. (2016) “Integrated Nutrient Management; Option for Improvement on Soil Chemical Properties, Growth and Yield of Cocoyam (Colocasia esculenta)”, Annual Research & Review in Biology, 10(4), pp. 1-12. doi: 10.9734/ARRB/2016/25739.

[5] Dasgupta, S., Sarkar, A., Chaitanya, A., Saha, A., Dey, A. and Mondal, R. (2017) “Response of Potato Crop to Integrated Nutrient Management in the Indo-Gangetic Alluvial Soils of West Bengal, India”, Journal of Experimental Agriculture International, 16(3), pp. 1-10. doi: 10.9734/JEAI/2017/33138.

Latest Research on Mineral Composition: Feb – 2020

The Distinction between Grain Size and Mineral Composition in Sedimentary-Rock Nomenclature

A system of grain-size nomenclature of terrigenous sediments and sedimentary rocks is introduced wherein fifteen major textural groups are defined on the ratios of gravel, sand, silt, and clay. Further subdivision of every class is predicated on the median diameter of every size fraction present. Next, the mineral composition of terrigenous sedimentary rocks is taken into account . A triangular diagram is employed to define eight rock types (orthoquartzite, arkose, graywacke, and five transitional types) supported the mineralogy of the silt-sand-gravel fraction and ignoring clay content. the author contends that the present practice of calling all clayey sandstones “graywackes” isn’t valid, inasmuch because it represents a confusion of texture with composition. [1]

Mineral composition and ash content of six major energy crops

The chemical composition of biofuels has not received adequate attention as long as it’s a crucial aspect within the introduction of energy crops. during this study, the ash content and mineral composition (C, N, Al, Ca, Cl, Fe, K, Mg, Na, P, S, Si) of stems, leaves and reproductive organs of some promising energy crops were determined and compared with the respective recommended thresholds reported in literature. Overall, cynara exhibited the very best ash and mineral contents, which indicate high slagging, fouling and corrosion tendencies. [2]

Mineral composition of non-conventional leafy vegetables

Six non-conventional leafy vegetables consumed largely by the agricultural populace of Nigeria were analyzed for mineral composition. Mineral contents seemed to be hooked in to the sort of vegetables. thorny amaranth and baobab leaves contained the very best level of iron (38.4 mg/100 g and 30.6 mg/100 g dw, respectively). These values are low compared to those for common Nigerian vegetables but above those for other food sources. All the vegetables contained high levels of calcium compared to common vegetables, thus they might be an upscale source of this mineral. Microelement content of the leaves varied appreciably. Zinc content was highest in Moringa oleifera, baobab and sicklepod leaves (25.5 mg/100 g, 22.4 mg/100 g and 20.9 mg/100 g dw, respectively) while the manganese content was comparatively higher in taro. [3]

Snow-dirt sludge as an indicator of environmental and sedimentation processes in the urban environment

The formation of snow-dirt sludge (SDS) via the blending of snow and surface-deposited sediment by vehicles and pedestrians is one among the smallest amount studied sedimentation processes in urban areas. The aim of this study was to analyse the fabric , mineral, particle size, and chemical compositions of the SDS solid phase. The study was conducted using Ekaterinburg, Russia, as an example city with an intense anthropogenic impact and an extended cold snowy period. the typical content of the SDS solid phase was approximately 35 g L−1 of meltwater on heavy traffic roads, of which the dust fraction. [4]

Phytochemical Properties, Proximate and Mineral Composition of Curcuma longa Linn. and Zingiber officinale Rosc.: A Comparative Study

Aims: to research the phytochemical property, proximate and mineral composition of turmeric and common ginger dried rhizomes.

Study Design: Activity directed phytochemical screening, proximate analysis and mineral composition investigation of C. longa and Z. officinale rhizomes using in vitro methods.

Place and Duration of Study: Medicinal Plants Section, Bioresources Development Centre, Ogbomoso, Nigeria between October and November, 2016.

Methodology: C. longa and Z. officinale rhizomes were separately washed dried (at room temperature) and pulverized. The powdered rhizomes were then used for phytochemical screening, proximate analysis and mineral composition investigation. [5]

Reference

[1] Folk, R.L., 1954. The distinction between grain size and mineral composition in sedimentary-rock nomenclature. The Journal of Geology, (Web Link)

[2] Monti, A., Di Virgilio, N. and Venturi, G., 2008. Mineral composition and ash content of six major energy crops. Biomass and Bioenergy, (Web Link)

[3] Barminas, J.T., Charles, M. and Emmanuel, D., 1998. Mineral composition of non-conventional leafy vegetables. Plant foods for human nutrition, (Web Link)

[4] Snow-dirt sludge as an indicator of environmental and sedimentation processes in the urban environment
Andrian Seleznev, Ilia Yarmoshenko, Georgy Malinovsky, Ekaterina Ilgasheva, Elena Baglaeva, Anastasia Ryanskaya, Daria Kiseleva & Tamara Gulyaeva
Scientific Reports, (Web Link)

[5] Taoheed, A. A., Tolulope, A. A., Saidu, A. B., Odewumi, O., Sunday, R. M. and Usman, M. (2017) “Phytochemical Properties, Proximate and Mineral Composition of Curcuma longa Linn. and Zingiber officinale Rosc.: A Comparative Study”, Journal of Scientific Research and Reports, (Web Link)

News Update on Proteins Research: Jan – 2020

Moonlighting proteins

The idea of 1 gene—one protein—one function has become too simple because increasing numbers of proteins are found to possess two or more different functions. The multiple functions of such moonlighting proteins add another dimension to cellular complexity and benefit cells in several ways. However, cells have had to develop sophisticated mechanisms for switching between the distinct functions of those proteins. [1]

Bone Morphogenetic Proteins

Bone morphogenetic proteins (BMPs) are multi-functional protein s that belong to the reworking growth factor β (TGFβ) superfamily. The roles of BMPs in embryonic development and cellular functions in postnatal and adult animals are extensively studied in recent years. Signal transduction studies have revealed that Smad1, 5 and eight are the immediate downstream molecules of BMP receptors and play a central role in BMP signal transduction. Studies from transgenic and knockout mice and from animals and humans with present mutations in BMPs and related genes have shown that BMP signaling plays critical roles in heart, neural and cartilage development. BMPs also play a crucial role in postnatal bone formation. BMP activities are regulated at different molecular levels. Preclinical and clinical studies have shown that BMP-2 are often utilized in various therapeutic interventions like bone defects, non-union fractures, fusion , osteoporosis and passage surgery. [2]

Intrinsically unstructured proteins

The recent suggestion that the classical structure–function paradigm should be extended to proteins and protein domains whose native and functional state is intrinsically unstructured has received an excellent deal of support. there’s ample evidence that the unstructured state, common to all or any living organisms, is important for basic cellular functions; thus it deserves to be recognized as a separate functional and structural category within the protein kingdom. during this review, recent findings are surveyed for instance that this novel but rapidly advancing field has reached some extent where these proteins are often comprehensively classified on the idea of structure and performance . [3]

Biotin proximity tagging favours unfolded proteins and enables the study of intrinsically disordered regions

Intrinsically Disordered Regions (IDRs) are enriched in disease-linked proteins known to possess multiple post-translational modifications, but there’s limited in vivo information about how locally unfolded protein regions contribute to biological functions. We reasoned that IDRs should be more accessible to targeted in vivo biotinylation than ordered protein regions, if they keep their flexibility in human cells. Indeed, we observed increased biotinylation density in predicted IDRs in several cellular compartments >20,000 biotin sites from four proximity proteomics studies. We show that during a biotin ‘painting’ time course experiment, biotinylation events in Escherichia coli ribosomes progress from unfolded and exposed regions at 10 s, to structured and fewer accessible regions after five minutes. [4]

Heat Shock Proteins in Cassia Species

Aim: Abiotic stresses, like drought, salinity, extreme temperatures, chemical toxicity and oxidative stress, are serious threats to plants. Heat shock proteins can assist in protein refolding under stress conditions at molecular levels. The proposed study was aimed to research and characterize the warmth shock proteins at the molecular level within the Cassia species.

Study Design: The experiments designed were intended to review and characterize the warmth shock proteins within the Cassia species using molecular approach.

Methodology: The samples were subjected to the varying heat treatments (30⁰C, 37⁰C and 42⁰C), and therefore the proteins obtained were further sequenced for similarity search. The peptide sequence was synthesized chemically and conjugated with Fissurella apertura hemocyanin (KLH) for the assembly of antibodies. Western blot was administered with the polyclonal antibodies to verify the results. [5]

Reference

[1] Jeffery, C.J., 1999. Moonlighting proteins. Trends in biochemical sciences, 24(1), (Web Link)

[2] Chen, D.I., Zhao, M. and Mundy, G.R., 2004. Bone morphogenetic proteins. Growth factors, 22(4), (Web Link)

[3] Tompa, P., 2002. Intrinsically unstructured proteins. Trends in biochemical sciences, 27(10), (Web Link)

[4] Biotin proximity tagging favours unfolded proteins and enables the study of intrinsically disordered regions
David-Paul Minde, Manasa Ramakrishna & Kathryn S. Lilley
Communications Biology volume 3, (Web Link)

[5] Pant, G., Kumari Chauhan, U., Malla, S., Banupriya, S. and Pati, R. (2017) “Heat Shock Proteins in Cassia Species”, Biotechnology Journal International, 17(4), (Web Link)