Effect of Aqueous Leaf Extract of Ocimum gratissimum on Antiretroviral Drug-Induced Hepatotoxicity in Rats | Chapter 13 | Recent Advances in Biological Research Vol. 4

Aim: Hepatotoxicity, among other adverse effects, constitutes one of the greatest impediments to successful antiretroviral drug therapy (ART) in HIV/AIDS patients. The main objective of the study was to determine if the aqueous leaf extract of Ocimum gratissimum has a protective effect on ART-induced hepatotoxicity in rats.

Place and Duration: Department of Pharmacology, Chukwuemeka Odumegwu University, Nigeria (six months duration).

Methodology: Twenty five (25) albino rats of both sexes were divided into 5 groups of 5 each and treated as follows: Group A (no antiretroviral drugs, no extract); group B (antiretroviral drugs alone); group C (extract alone); group D (antiretroviral drug plus 40 mg/kg extract); group E (antiretroviral drug plus 80 mg/kg extract). All treatment lasted for twenty eight days. Blood samples were collected and serum alanine transaminase (ALT) and aspartate transaminase (AST) determined using UV-spectrophotometer. Thereafter, the animals were sacrificed and their livers harvested and examined histologically. The mean (± S.E.M) of data were calculated and further analyzed for statistical significance using graph Pad Prism 5.0.

Results: Mean serum ALT were 35.6±6.4, 54.0±9.4, 53.8±22.9, 90.5±21.9, 86.5±13.9 and that of AST were 143.8±19.7, 205.2±14.9, 58.0±27.9, 162.3±41.4, 150.5±44.8 for groups A, B, C, D, and E respectively. There was a statistically significant difference between the mean values of serum AST for group B and those for group C (p value of 0.016). However there was no statistically significant difference between the ALT values for the test and control groups of rats (p value > 0.999). Also, there was no statistically significant difference between the mean values of  AST for group B and those of groups A,  D, E (p value = 0.659). The histology report for the liver was normal for all groups.

Conclusions: This extract did not produce significant reduction of serum ALT and AST in ART-treated rats in this study However, this reduction in serum aminotransferases was not observed in the groups that received antiretroviral drugs plus the extract. These statistical findings show that there was no statistically significant difference in serum ALT and AST between the exposed and control groups (p > 0.05). Consequently, the null hypothesis was accepted and the alternative hypothesis rejected. Therefore, it could be concluded that at the doses of ART and extract and length of exposure used in this study, aqueous leaf extract of Ocimum gratissimum did not reduce the serum level of ALT and AST in rats.

Author(s) Details

Associate Professor Peter Ughachukwu
Department of Pharmacology and Therapeutics, College of Medicine, Chukwuemeka Odumegwu Ojukwu University, Nigeria.

Emeritus Professor Paul Okonkwo
Department of Pharmacology and Therapeutics, College of Medicine, Chukwuemeka Odumegwu Ojukwu University, Nigeria.

Dr. Cornelius Nwozor
Department of Human Physiology, College of Medicine, Chukwuemeka Odumegwu Ojukwu University, Nigeria.

Dr. Joseph Okafor
Department of Anatomy, College of Medicine, Chukwuemeka Odumegwu Ojukwu University, Nigeria.

Dr. Mrs. Ifeyinwa Nwafia
Department of Medical Microbiology, College of Medicine, Chukwuemeka Odumegwu Ojukwu University, Nigeria.

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Variation of Mineral Micronutrient Elements in Robusta Coffee (Coffea canephora Pierre ex A. Froehner) As Measured by Energy Dispersive X -Ray Fluorescence | Chapter 12 | Recent Advances in Biological Research Vol. 4

Introduction: In many parts of the world, Coffea grows between latitude 25°N and 25°S and requires specific climatic conditions to produce high bean yield and quality. In Uganda, Robusta coffee is grown in a diversity of soils that range from red sandy clay or gravelly loam to soft laterite. Macro-elements such as K, Ca and Na though detectable through EDXRF are not as heavy as trace elements such as Fe, Zn, Cu, Mn, a factor that makes the micro-elements more amenable to EDXRF measurement. The EDXRF spectrometer instrument has a semi-conductor detector that measures the entire energy spectrum when placed closer to the sample to minimize loss of energy by the fluorescent X-ray.

Aim: Robusta coffee growing countries of the world such as Uganda, have experienced a decline in market prices since 2002 due to changed consumer preferences for other beverages. In Uganda, where Robusta coffee is the major income earner, one practical way of ensuring that coffee prices remain competitive is to enhance quality. The aim of this study was to analyze the extent of macro and microelement densities in the soils of Robusta coffee growing Ugandan farms, their relationships with density levels in the leaf and in the green bean and their ultimate contribution in the determination of cupping and marketing qualities. This information would enable coffee producers in Uganda to make decisions on which organic and inorganic fertilizer applications are needed to alleviate coffee quality which in turn is the key determinant of world prices.

Study Design: Soil, leaf and ripe cherry samples were derived from sixty seven tagged trees in twenty three on farm locations at Kawanda and Entebbe in fields that had no fertilizer application history.

Place and Duration of the Study: This work was conducted at the National Agricultural Coffee Research Institute (NaCORRI), Uganda and the Institute of Nuclear Science and Technology, University of Nairobi, Kenya, between January 2010 and December 2013.

Methodology: Nutrient densities of potassium (K), calcium (Ca), iron (Fe), titanium (Ti), zinc (Zn), copper (Cu), boron (B), lead (Pb) and manganese (Mn) of sixty seven soil, leaf and green bean samples collected from 23 districts were measured with Energy dispersive x-ray fluorescence (EDXRF). All the samples were oven-dried at 800C for 18-20 hours and ground to about 50 mm sieve-size. Three pellets of 2.5 cm in diameter were made and irradiated with a Cd-109 radioactive source. The X-rays emitted by the elements were detected by liquid nitrogen cooled Si (Li) detector and data analyzed with the Quantitative X-ray Analysis System (XQAS/AXIL) software.

Results: The results showed that the mean concentrations of Ca, Mn, Fe and Zn were highest in the soil but lowest in green beans. K concentrations were higher in green beans than in the leaf and were lowest in the soil. In most cases, the concentrations of microelements found in the soil determined the levels in the leaf and in the green bean. Microelement availability and concentrations were influenced by the interaction between the various elements, tree age, elevation, soil texture and location.

Conclusions: Mineral element K and microelement Cu were found in their highest concentration in the green bean. Mn, Zn and B were important elements in the determination of organoleptic cup attributes whereas soil, leaf and green bean nutrient concentrations were important in determining the medium size of the bean. EDXRF is an effective tool in the measurement of micronutrient concentrations of Robusta coffee growing soils, leaves and green beans. Cu was an essential microelement of physiological and metabolic processes in both leaves and in the green bean whereas Mn, Zn and Fe are critical microelements during photosynthesis. While all the trace elements measured, namely, Mn, Cu, Fe and Zn were crucial determining factors in bean and cup qualities of Robusta coffee, Zn and Mn were the more important microelements during fruit development. Soil Mn positively influenced flavor and aroma cup attributes whereas green bean amounts of B and Zn positively influenced trigonelline and caffeine concentrations.

Author(s) Details

Prof.  Kahiu Ngugi
Department of Plant Science and Crop Protection, Faculty of Agriculture, College of Agriculture and Veterinary Sciences, University of Nairobi, P.O.BOX 29053-00625, Kangemi, Nairobi, Kenya.

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Total Antioxidants Status, Selenium Concentration, Catalase and Glutathione Peroxidase Activities in Rabbits Fed a High-Garlic Diet | Chapter 11 | Recent Advances in Biological Research Vol. 4

Aim: Results from previous studies on the effects of intake of garlic on the body antioxidant system appear inconclusive. This study measured glutathione peroxidase (GPx) and catalase (CAT) activities, Total Antioxidant Status (TAS) and selenium (Se) level in rabbits fed a high-garlic diet.

Methods: Ten rabbits randomly assigned into two groups (group 1=control and group 2= test) of five rabbits each were used for the experiment. The diet of group 2 was mixed with raw garlic homogenate (1.75g/Kg body weight/day) for 4 weeks. Two sets of blood samples were collected from each animal at the beginning of the experiment (baseline) and at the end (4 weeks). Blood samples were analyzed for blood GPx and serum CAT activities and TAS by spectrophotometric methods while Se was determined using Atomic Absorption Spectrophotometric (AAS) method.

Results: At baseline, means of the parameters measured did not differ significantly between groups 1 and 2. At the 4th week, means of TAS (1741.20±381.53 µmol Trolox equiv/L), Se (47.20±12.22 µg/dl), CAT activity (273.20±68.05 U/L) and GPx activity (12392.00±3068.34 U/L) in the test group were significantly higher than means of the corresponding control group (820.20±91.94 µmol Trolox equiv/L, 20.80±1.92 µg/dl, 145.40±13.35 U/L, 8528.00±1757.59 U/L, respectively). There were positive associations between CAT and GPx (r = 0.65, p<0.01) activities, CAT and TAS (r = 0.77, p<0.01), GPx and TAS (r = 0.66, p<0.01) and Se and TAS (r = 0.70, p<0.01).

Conclusion: Results from this present study suggest that significant increases observed in the activities of antioxidant enzymes, Se level and TAS could possibly be associated with consumption of high garlic diet by the rabbits. Overall, the significant increase in the CAT and GPx activities as well as the levels of Se and TAS seen in the test group at the end of the study suggests that consumption of raw garlic extract could result to increases in the activities of important antioxidant enzymes and improve the total antioxidant status of the body of rabbits.

Author(s) Details

Japhet M. Olisekodiaka
Department of Chemical Pathology, College of Health Sciences, Nnamdi Azikiwe University, Awka, Nigeria.

Christopher Igbeneghu
Department of Biomedical Sciences, College of Health Sciences, Ladoke Akintola University of Technology, Ogbomoso, Nigeria.

Adekunle A. Adesiyan
Department of Biomedical Sciences, College of Health Sciences, Ladoke Akintola University of Technology, Ogbomoso, Nigeria.

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Induction of Mitochondrial-Mediated Apoptosis by Solvent Fractions of Methanol Extract of Heliotropium indicum in Rat Liver Cells | Chapter 10 | Recent Advances in Biological Research Vol. 4

Introduction: The biochemical process of apoptosis may occur by a number of mechanism including; extrinsic or death-receptor pathway, intrinsic or mitochondrial-mediated pathway which involves the opening of Mitochondrial Membrane Permeability Transition (MMPT) pore in situations of intracellular calcium overload or oxidative stress or cellular insult and by the perforin/granzyme pathway. There is considerable evidence that bioactive agents in some plants (sulforaphane in cruciferous vegetables, genistein in soybeans and epigallocatechin gallate in green tea etc.) were able to induce MMPT pore opening. Quite a number of these agents have been identified and they currently under preclinical and clinical trials.

Aim: Mitochondrial Membrane Permeability Transition (MMPT) pore has emerged as a promising target for drug development because the release of cytochrome c upon the opening of the pore is a point of no return for mitochondrial-mediated apoptosis to occur. Heliotropium indicum (HI) is as an anti-tumor and wound healing agent in traditional medicine. It is not known whether its mode of action involves the induction of apoptosis via the opening of the MMPT pore.

Methodology: Mitochondria, isolated from male albino rat liver (about 100 g), were exposed to varying concentrations (10, 30, 50, 70, and 90 µg/ml) of  solvent fractions of  methanol extract of HI  i.e  Chloroform (CFHI), Ethylacetate (EFHI), Methanol (MFHI)  and crude  Methanol Extract (MEHI) of HI. Opening of the pore, cytochrome c release, mitochondrial ATPase activity and extent of mitochondrial lipid peroxidation were assessed spectrophotometrically in vitro. Activation of caspases 9 and 3 were also assessed using ELISA kits.

Results: In the absence of Ca2+, CFHI, EFHI, MFHI and MEHI induced the opening of the pore in a concentration-dependent manner with CFHI having the highest induction fold of 26 and MFHI as the lowest having 6.6. All the fractions inhibited lipid peroxidation in a concentration-dependent manner. Also, these fractions induced the release of cytochrome c with CFHI having the highest effect and the least by MFHI. Mitochondrial ATPase activity was enhanced by all the fractions with CFHI having the highest stimulatory effect. Interestingly, intra-peritoneal administration of CFHI and MEHI at 2, 5, 10 and 20 mg/kg body weight for 21 days resulted in significant opening of the pore, the release of cytochrome c and activation of caspases 9 and 3. All these effects were highest with 20 mg/kg body weight.

Conclusion: These findings therefore suggest that Chloroform Fraction of Heliotropium indicum is the most potent of all these fractions and therefore contains the bioactive agent that induces mitochondrial-mediated apoptosis in normal liver cells. The fraction will therefore be useful for further studies for drug development in diseases requesting up-regulation of apoptosis.

Author(s) Details

Adeola O. Olowofolahan
Laboratories for Biomembrane Research and Biotechnology, Department of Biochemistry, College of Medicine, University of Ibadan, Nigeria.

Yemisi D. Adeoye
Laboratories for Biomembrane Research and Biotechnology, Department of Biochemistry, College of Medicine, University of Ibadan, Nigeria.

Olufunso O. Olorunsogo
Laboratories for Biomembrane Research and Biotechnology, Department of Biochemistry, College of Medicine, University of Ibadan, Nigeria.

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Castanea sativa Mill: A Brief Review | Chapter 09 | Recent Advances in Biological Research Vol. 4

The use of medicinal plants has grown all over the world. Nowadays the ethnopharmacological applications of medicinal plants are an important matter when it comes to search for new molecules with biological activity.

Castanea sativa Mill is a species with great economic value since chestnut trees are an important crop in the northern region of Portugal and new applications of this plant are being developed by the pharmaceutical and food industries. Beyond its economic importance it also has beneficial effects in human health as consequence of their biological activities namely antioxidant, antimicrobial, antispasmodic, among others.

In this work we present a review of the main bioactive compounds that constitute the chestnut extracts, their toxicity and their main biological activities.

Author(s) Details

José Manuel Neves
Faculty of Health Sciences, University Fernando Pessoa, Rua Carlos da Maia, 296 4200-150 Porto, Portugal.

Sílvia Cunha
Faculty of Health Sciences, University Fernando Pessoa, Rua Carlos da Maia, 296 4200-150 Porto, Portugal.

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Estimation of Stability of Arterial Pressure and Pulse at Changes of Geomagnetic Data and Atmospheric Pressure | Chapter 08 | Recent Advances in Biological Research Vol. 4

This work analyses annual blocks of daily observations of pulse with arterial blood pressure (ABP) and interrelation between them. Dynamics of morning and evening parameters is compared, behavior of atmospheric pressure and geomagnetic activity is correlated. Pulse and arterial blood pressure are evaluated during periods of more active behavior of patient (expeditions, business trips, holidays, etc), as well as during moments of considerable deviations of atmospheric pressure from annual average value. Regular (in the morning after sleep and in the evening) observations of heart rate (HR), systolic arterial blood pressure (SABP) and diastolic arterial blood pressure (DABP) are used. These observations are conducted by the patient (man with 2 step and 2 stage hypertensive disease, 1940 year of birth) within the program of ABP self-control. Values of geomagnetic activity and atmospheric pressure, drawn in for correlation, are tied to patient’s place of residence (Troitsk, Moscow). This study is based upon series reflecting in time dynamics of correlation coefficients, which were received during simultaneous scanning of characteristics under study. Such series of correlation coefficients of medical parameters, analogue of degree of organism coordination are taken as the basis and compared with changes of environmental parameters. The suggested approach made it possible to single out situations, at which interrelations of pulse and ABP may be compared with concrete environmental conditions. A number of differences in characteristics of morning and evening series of monitoring may be tied with rapid relaxation of organism during night from rhythmical activity and enhanced sensitivity of organism in the morning to background effect. The considered material has confirmed effectiveness of self-control of ABP for its control and during selection, if necessary, of antihypertensive drug, which ensures normal patient’s vital activity.

Author(s) Details

Igor Shibaev
Pushkov Institute of Terrestrial Magnetism, Ionosphere and Radio Wave Propagation (IZMIRAN), Troitsk, Moscow 142190, Russia.

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Bioconversion of Sweet Potato Leaves to Animal Feed | Chapter 07 | Recent Advances in Biological Research Vol. 4

Background: High cost of conventional animal feed ingredients in Nigeria has made it necessary to search for alternative local sources of feed. Crop residues including sweet potato leaves abound in Nigeria. These have been explored as feed sources. The ability of microorganisms to convert agricultural wastes to more useful products could be harnessed to produce feed from sweet potato leaves which can be obtained in high abundance at low cost.

Aim: To examine the possibility of converting sweet potato leaves to animal feed through fermentation with a co-culture of Chaetomium globosum and Saccharomyces cerevisiae.

Materials and Methods: Triplicate samples of sweet potato leaves were fermented with a co-culture of C. globosum and S. cerevisiae for 21 days at 25±2°C and the effects of fermentation on nutrient composition was determined. Fermentation and control samples were analysed for proximate, amino acids, and elemental contents. Acute oral toxicity of the fermented leaves was determined by the fixed dose method using mice and rats. Feed value of the fermented sweet potato leaves for mice was determined. Parameters assessed included feed intake, protein intake, weight gains, feed efficiency ratio, and protein efficiency ratio.

Results: Crude protein, crude fat and ash contents increased by 97.5%, 265.3% and 12.3%, respectively, while crude fibre and nitrogen free extract values decreased by 22.7% and 61.4% respectively. Energy content increased by 14.5%. The observed changes in the values of these nutritional components were significant (P < .05). The percentage dry matter values of all the amino acids analyzed (lysine, histidine, arginine, aspartic acid, threonine, glutamic acid, proline, glycine, alanine, cystine, valine, methionine, isoleucine, leucine tyrosine and phenylalanine) were found to increase, with the contents of seven of the amino acids increasing significantly. Calcium, phosphorus, potassium and magnesium contents increased significantly while those of copper and iron decreased. The fermented leaves were found to be non toxic to mice and rats. Sole use of fermented sweet potato leaves by the mice led to depression in feed intake, weight gain, feed efficiency ratio and protein efficiency ratio. Mice fed with commercial mice feed supplemented  with 5% fermented sweet potato leaves had higher weight gains, feed efficiency and protein efficiency ratios than those fed on the commercial mice feed.

Conclusion: Fermentation of sweet potato leaves with a co-culture of C. globosum and S. cerevisiae improved the nutritional value of the leaves. Fermented sweet potato leaves can be included in mice feed up to 5% without negative effects. Sole use of fermented sweet potato leaves as feed for mice and possibly other animals would require mineral supplementation, energy enhancement, and further crude fibre reduction.

Author(s) Details

Isaac A. Onyimba
Department of Science Laboratory Technology, University of Jos, P.M.B. 2084, Jos, Nigeria.

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Temporal Variations in the Short and Long Term of the Species Richness and the Abundances Structuring in a Same Butterfly Community: An Enlightening Case Study at Gariwang-san (Korea) | Chapter 06 | Recent Advances in Biological Research Vol. 4

The way species-rich communities react over time, in both the short term (inter-annual) and the long term (decades) is usually very complicated but also often proves very instructive as regards several aspects of the functional structuration of the community. A three decades monitoring of the same butterfly community at Gariwang-san (South-Korea) thus provides interesting insights on the temporal responses of several major features of the community structuration. In turn, the close analysis of these responses allows to relevantly address some hotly debated hypothesis. The two following ones are addressed and questioned in particular: (i) which kinds of structural and functional features of communities prove being more or less sensitive to environmental degradations and (ii) to what extent the notions of “functional equivalence” and “community resilience” are likely supported by the analysis derived from this short-term and long-term monitoring.

Besides, on a methodological point of view, it is especially emphasized the absolute necessity, in this type of studies, to consider (sub-) exhaustive samplings of the targeted communities, since incomplete inventories would most often lead to severe bias in the interpretation of results. Alternatively, if (sub-) exhaustive samplings cannot be achieved (as is often the case in practice), it is mandatory to complete the available partial samplings (and the associated incomplete species abundance distributions) by an appropriate procedure of numerical extrapolation.

Author(s) Details

Dr. Jean Béguinot
Biogéosciences, UMR 6282, CNRS, Université Bourgogne Franche-Comté, 6, Boulevard Gabriel, 21000 Dijon, France.

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Phytochemical and Antiepileptic Activity of the Ethanol Leaf Extracts of Culcasia falcifolia in Pentylenetetrazole Induced Seizure in Mice | Chapter 05 | Recent Advances in Biological Research Vol. 4

Aim: To investigate the phytochemicals and the anticonvulsant activity of the ethanol leaf extract of Culcasia falcifolia used in the traditional medical treatment of epilepsy.

Methodology: The phytochemical screening was carried out using standard protocol while the anticonvulsant activity was studied using Pentylenetetrazole in mice.

Result: The preliminary phytochemical screening carried out on the ethanol extract of Culcasia falcifolia revealed the presence of alkaloids, flavonoids, saponins, tannins, polyphenols, and glycosides. In the anticonvulsant activity, there was a significant (*p< 0.05, **p<0.01) increase in the mean latency of tonic convulsion (243.72 ± 6.90*, 402.56 ± 5.52**) and significant (*p< 0.05, **p<0.01) decrease in the mean duration of tonic convulsion (192.62 ± 7.72*, 158.99 ± 8.66**) in a dose-dependent manner at the dose of 200 and 400 mg/kg body wt. respectively. The extract at 400 mg/kg body wt. showed 100% protection against mortality.

Conclusion: The results of this study suggest that the ethanol leaf extract of Culcasia falcifolia possesses anticonvulsant activity in PTZ induced seizure in mice. Therefore, the findings of the present study, suggests that ethanol extract of Culcasia falcifolia leaves has anticonvulsant activity against PTZ-induced seizures in mice by modulating GABA receptor-mediated inhibitory neurotransmission, reducing the T-type of Ca++ currents, activating GABAA receptors and facilitating the GABA-mediated opening of Chloride channel. Based on the folkloric claims, we infer that the decoction of the leaves of Culcasia falcifolia could be integrated into health care system for the management of epilepsy.

Author(s) Details

A. Gracelyn Portia
Department of Biological Sciences, University of Eastern Africa, Baraton, P.O.Box – 2500, Eldoret – 30100, Kenya.

D. M. Mwaniki
Department of Biological Sciences, University of Eldoret, P.O.Box 1125-30100 Eldoret, Kenya.

A. G. M. Ng’wena
Department of Medical Physiology, Maseno University, Maseno – 40105, Kenya.

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Natural Compounds in the Treatment of Inflammatory lung Diseases: An Approach on Eucalyptol | Chapter 04 | Recent Advances in Biological Research Vol. 4

It is estimated that there will be an increase in the incidence of chronic obstructive pulmonary disease (COPD) in the coming decades. Thus, the pharmacological attributes of products of plant origin should be considered as an important economic and scientific strategy in the investigation of therapeutic alternatives, since their experimental validations are indispensable to substantiate the reliability of these products in the treatment of chronic diseases. Like biologically active compounds, Eucalyptol, also known as 1,8- cineole, is the major constituent of the leaf oil of eucalyptus species, such as Eucalyptus globulus and Eucalyptus tereticornis. It is a terpenoid oxide, free of steroid-like side effects. This study is based on a review of the specialised literature with purpose to discuss the biological effects of Eucalyptol in the respiratory system and its interaction with some of the most promising targets in the treatment of COPD, such as: receivers and membrane channels, oxidative stress, transcription and expression of cytokines, cell adhesion molecules and neutrophil chemotaxis, proteases and remodeling.

Author(s) Details

Fladimir de Lima Gondim
Postgraduate Program in Physiological Sciences, State University of Ceará, Ceará, Brazil.

Gilvan Ribeiro dos Santos
Center of Technological Sciences, State University of Ceará, Ceará, Brazil.

Daniel Silveira Serra
Center of Technological Sciences, State University of Ceará, Ceará, Brazil.

Francisco Sales Ávila Cavalcante
Center of Technological Sciences, State University of Ceará, Ceará, Brazil.

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