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I'm Sarah, a twenty something book hoarder and travel lover…
…sharing real & fictional escapism. I'm particularly passionate about being as authentic as possible in all my writing, whether that's travel tips, book reviews or just general life chat.
I've always found a way to travel whilst working full time, whether that was cheap flights and 48 hours in Copenhagen or squeezing in a trip to Bolivia AND Peru into a two week holiday. However in 2019, I was fortunate enough to take time off work and travel South East Asia for a few months, which was literally a dream come true. Since my return I've been living up North in the UK and trying to find my feet again in the "real world".
When I'm not blogging you can catch me reading, drinking coffee, looking for sustainable products to use or pushing my bodily limits in YouTube yoga videos of cute girls on beaches (wishing it was me).
Note: Despite the circumstances I'm continuing to share my travel content from my trip last year. We don't know how the future of travel looks but we can all dream about destinations we'd like to visit in the future.
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God's Plan for Pregnancy Support Meetings – 2016 Come along and discover God's Plan for your Conception, Pregnancy and Childbirth!
Tag Archives: Prayer
Conception Prayer
I am always receiving requests from couples asking for prayer for healing of infertility and while I would love to pray for everyone, time does not permit. Therefore, I have recorded a prayer that the Lord laid on my heart to help you.
Please do not view this prayer as a quick fix or something to replace your own prayers but rather as a point of contact for you to connect your faith with. I believe the Lord will anoint this prayer so that it will help you to experience the breakthrough you need in your body to see your miracle baby come into manifestation.
And when you do I would love to hear of your testimony!
To view the prayer visit: http://www.youtube.com/watch?v=-moXkuxVIZU&feature=share&list=UUKlfS2XAJYfdvFqHY9nnLtQ

subset: commoncrawl


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Cricket in South Africa is synonymous with the hot African summer. In fact, nothing says that the summer has arrived like sitting on the grass embankment under the oaks during
England rout means there will be no quick fix for Proteas
The organisers of IPL 2019 really could not have wished for a better end to the tournament. Sunrisers Hyderabad aside, these were arguably the two best teams in the competition
One of the greatest cricketers of all time, Shane Warne, is also known to be a keen gambler and poker player. There could be a reason why the Australian legend
The world of English cricket has seen many marvellous talents over the years but, following his incredible test-winning innings at
Jofra Archer's inclusion in the England World Cup squad has been incredibly fast-tracked after making his debut in May 2019
This year's Cricket World Cup, the 12th edition of this tournament, is scheduled between May 30 and July 14. The
Virat Kohli A mixed bag for Kohli leading into the World Cup. India's form heading into the competition hasn't been
The 2019 ICC World Cup is set to be hosted by England and Wales from May 30 to July 14.
Former England captain Kevin Pietersen said that India's senior off-spinner Ravichandran Ashwin will not get much support for the controversial


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"It's November 10th, feeling like fall with this 40-50 degree weather. I survived Halloween and was able to reject 'most' of the candy that came my way. Mainly because I was conscious of my plan to change. I was able to listen to a podcast from Greg Gassman (founder of CrossFit) and Chris Cooper recently and Greg had so many points that stood out to me. I am going to highlight and elaborate on some of those points.
Greg states he would rather 'you eat right and don't exercise than exercise and eat like $#!@'. This is so true. Why put in all the effort to exercise and not back it up with proper diet. The old saying 'Abs are made in the kitchen' holds so much meaning. Often when patients are trying to tell me how they are implementing lifestyle changes, they tend to point out how they are walking more, exercising more and being more active, but all that does not matter if you continue to consume processed foods and sodas. The exercising is the easy part, and we Americans are looking for fast and easy.
Greg also points out that there is no '3rd element'. We have nutrition and exercise and that's it! However, we tend to keep searching for more, such as, supplements, the magic pill, or things like ice baths. We feel the need to want to rely on 'something else' to help us get healthy and perform better instead of what really matters – what you eat and how you move!!! This is basic stuff people, but yet not easy (or at least perceived as not easy). I am not against supplements, such as fish oil etc, however, until you have tried the basics of nutrition to control cholesterol and triglycerides, I agree with Greg, you shouldn't be adding a '3rd element'.
Greg also mentioned 'non-responders'. This was a term that actually made me laugh and one that I will be using from now on. You see NO results. This happens over and over again in the when people are trying to get fit. They never align the fitness and nutrition and don't see the results they are looking for. They often get frustrated and ask for a '3rd element'. Maybe a pill, supplement, or something they saw on TV from Dr. Oz. They are searching for a quick answer, yet the answer has been given to them over and over and over. Cut out the processed foods, quit drinking sodas (including diet drinks) and move more!!!
I feel at this time the best way to get people to change is to hold them accountable, show you care and follow through. Greg mentioned to 'have them write it down'. This offers accountability. Log your meals, keeping track helps with awareness and accountability. I would even go further and say, as a doctor, meet your patients at the grocery store instead of the doctor's office. Demonstrate 'how to squat' (or get out of a chair) in the office. These are things patients need to know. They are already aware they need to change and eat better, so knowing what one needs to do (and education) is not the missing piece. I feel the missing piece is the accountability.
This accountability can be found in many different forms. It may be a friend, a spouse, a gym, a doctor (rarely), a nutrition program or even an app on their iPhone. They set goals, the measure their goals, and they have someone looking over them to ensure they are meeting their goals, and/or redirecting them continuously.
So I am challenging YOU and MYSELF to find the accountability that works for you. Stop looking for the '3rd element' to get you healthy and fit. Go back to the basics, eat what is natural and not processed. Learn to move, and move correctly and often. Develop some variability in order to keep from getting bored, and when you fail, pick right back up and start over. WE CAN DO THIS AND WE MUST!


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Filed under: Obsessions | Tags: Courtney Cox, David Arquette, gold necklaces, jewelry, Obsession, Satya Jewelry, Yoga
Yoga has been around forever, but only recently has yoga inspired fashion. I've been experimenting with yoga for about two years now. I love the serenity and calmness that comes with the practice. Jewelry designers have utilized the symbols for peace within yoga (like the Om symbol, the tree of life, the lotus etc) and made gorgeous jewelry designs. One of my favorite designers is Satya Jewelry. Satya means "truth." The company was started by designers Satya and Beth. They combine spiritual symbols, healing gemstones and the principles of yoga into their creations. Also, a percentage of their proceeds goes to various childrens' organizations through the Satya foundation. The jewelry is so gorgeous! Celebrities such as Rachel Bilson and Courtney Cox have worn their designs. I can't wait to get one of their gold necklaces!
Source: http://meetthefamous.com/celebrity-closet/, http://www.satyajewelry.com/


text: sskendre 0 Report post Posted October 7, 2015 Dear All, I am not able to downlaod Barcode Genie User Guideand other data which is available on link http://papilio.cc/index.php?n=Papilio.BarcodeGenie Please help me or share data if it is available. Thanking you, With regards,S. S. Kendre Share this post Link to post Share on other sites


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The MANY Uses of Vinegar
The many uses of vinegar is quite an impressive list! While vinegar has long been used as a natural cleaning, disinfecting and beauty aid, did you know it could do all of this? Of course, always test a small area first before trying any of these remedies and be sure to consult a doctor when using it as a health aid!
-To wash no-wax floors, add ½ cup of white distilled vinegar to a half-gallon of warm water.
-To clean windows without streaking, mix equal parts of white distilled vinegar and warm water. Dry with a soft cloth.
-To remove water rings on wood furniture, rub a mixture of equal parts of white distilled vinegar and olive oil. Rub with the grain and polish for the best results.
-Rub down shower doors with a sponge soaked in white distilled vinegar to remove soap residue.
-Prevent icing on car windows by wiping the windows the night before with a solution of one part water to three parts white distilled vinegar.
-Deodorant and antiperspirant stains may be removed from clothing by lightly rubbing with white distilled vinegar and laundering as usual.
-Kill bacteria AND soften your clothes by adding 1 cup of white vinegar to your rinse cycle. No more need for fabric softener!
-Clean that hummingbird feeder regularly with equal parts apple cider vinegar and hot water. Rinse with cold water and air dry before re-filling with food.
-Add 2 tablespoons white distilled vinegar to 1 pint water and use to wash fresh fruits and vegetables, then rinse thoroughly. Research has shown that vinegar helps kill bacteria on fruits and vegetables.
-Prevent eggs from cracking when boiling them by adding 2 tbsp of vinegar to the water before boiling the eggs.
-Improve the flavor of boiled ham by adding 1 tablespoon vinegar to the cooking water.
-Not sure your baking soda is still good? Pour 2 tablespoons vinegar in a small dish, and add 1 teaspoon baking soda. Good baking soda should make the vinegar froth significantly.
-Deodorize your child's stinky lunchbox by soaking a slice of bread in white vinegar and then leaving it in your child's lunchbox overnight. Should be odor-free by morning!
-Did you leave that produce out just a bit too long and now have fruit flies/gnats?? First remove the source of the problem by throwing away the over-ripened produce and then place a bowl filled with ½ quart water, 2 Tbsp. sugar, 2 Tbsp. apple cider vinegar and a couple of drops of dish soap on the counter to attract the fruit flies.
-To keep your garbage disposal fresh with vinegar cubes. Make your own vinegar cubes by filling an ice tray with a mixture of 1 cup of vinegar and enough water to fill the ice tray and then freezing it. Run the mixture through the disposal, and then flush it with cold water for one minute.
-Rid your refrigerator and freezer of bad odors by cleaning the insides with a solution of equal parts vinegar and water, then wiping dry.
-Freshen your dishwasher by filling the soap reservoir with white vinegar and run the machine through a wash cycle.
-Clean your electric can opener blades with a toothbrush and white vinegar. Dip the toothbrush in white vinegar and position the bristles against the around the edge of the wheel. Turn on your can opener and let the brush do the work.
-Add one to two capfuls of apple cider vinegar to your warm bath. It draws toxins out of the body, leaving behind toned and moisturized skin. (also great to soothe sore muscles)
-Soothe sunburn pain by soaking washcloths in apple cider vinegar and laying them on the affected area.
-Soak your fingertips for 1 minute in 2 teaspoons vinegar and 1/2 cup warm water before applying the polish to make your manicure last longer.
-Pour one ounce of vinegar in your hair and rinse after shampooing to leave it soft and shiny.
-Apply a small amount of vinegar to bug bites to relieve itching.
What our many uses of vinegar do you and your family use?
Household Repurposing Ideas: 5 Clever Uses for Vinegar


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While the year is drawing towards its final quarter, those eager for new music still have plenty of things to be excited about. Whether you're into hip-hop, post-hardcore or punk, there are still great things to be heard in coming weeks for those whose most anticipated album has yet to see the light of day. Regardless, here are three tracks from albums not quite in our hands that have me excited to hear the final product when it hits the streets in the coming weeks.
1. "Marvin's Room"- Drake
Drake is kind of the Jekyll and Hyde of hip-hop. On one side, we have a clever wordsmith who has shown he can flow with some of the biggest names in the rap game, while on the other side we find Drake about to pour out soulful melodies take us further into the Rn'B realm than anything else. And while the other tracks surfacing from Take Care find Drake spitting more than swooning, "Marvin's Room" is an emotional, albeit blunt ride through getting someone back. Though many prefer Drake to rap instead of sing, count at least one person who wouldn't mind if Drake kept doing songs like this one, as the beat and lyrics make it a solid mix for a song meant to stir the emotional pot.
2. "Edward Benz, 27 Times" – La Dispute
Topping the emotionally epic Somewhere at the Bottom of the River Between Vega and Altair might be tough in the eyes of the droves of fans this band has won over since its release, but after a couple solid tracks surfaced through the past couple weeks, this track appeared through my Tumblr this afternoon. Possibly heavier lyrically than anything the band has penned before, the story of a man and his son is told through fits of twang-lined jamming guitars and a steady rhythmic attack. The track progresses with the story, adding bits and pieces musically as the band weaves through subtle melodies and strong drumming. Jordan Dreyer's emotional vocals pair with the wandering arrangements as the track apexes with his shouts of 'That's all I can offer, that's all that I know how to give.' Showing their growth in songwriting maturity since Somewhere, this track is lyrically and musically a seamless, natural progression for the band.
3 "Black Mass" – This is Hell
Though This is Hell released Weight of the World last year, Travis Reilly and company sound rejuvenated in the title track to their upcoming LP by the same name. After a simple, haunting opening, the track shreds through double-bass laced riffing that seemingly never lets up until Reilly's howl hails the verse into your ears. Armed with sludgy riffs and a chorus based around the haunting opening riff (all topped off with a gong!), the track pairs gritty metal with the band's punk undertones to hint at this record being a contender for hardcore album of the year.


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Location: McLeod Suites, 3rd floor (Curry Student Center)
Time: 8:30am - 12:30pm
Topics: StrengthsFinder and Free Assessment Resources
There seems to be a lot of talk about strengths in career counseling circles and many counselors have committed to formal training in order to become a Gallup Certified CliftonStrengths Coach. If you're considering this certification or just want to learn more about StrengthsFinder and its uses, this presentation by Anne Grieves, a Gallup Certified CliftonStrengths Coach will be of interest to you.
Anne will discuss how this tool is used specifically in career counseling, how it differs from the MBTI, and why so many coaches are embracing it. The presentation will be interactive and will include an activity to assess the alignment between your strengths and your work as well as a discussion of career counseling case studies.
Speaker Bios
Anne Grieves is an Associate Director in Career Development at Northeastern University as well as a private career coach/team building consultant. She received her Master's in Higher Education Administration from Teacher's College, Columbia University and is a certified Gallup Clifton Strengths Coach. Anne enjoys guiding individuals, groups and teams towards identifying and achieving their goals. Collaborating with Anne, her clients learn how to focus on their individual strengths in order to maximize their personal and professional potential. Anne's unique approach provides people with the tools they need to experience a paradigm shift in how to explore new possibilities, and re-imagine their goal-setting. Her two daughters, husband and dog fill the unscheduled hours in her day.
8:30am Registration/Breakfast
9:00am Announcements
9:15am Needs and Leads
9:30am StrengthsFinder + Q&A (Anne Greaves)
10:30am Break/Networking
11:00am Free Assessments
11:55am Wrap Up
We plan to feature speakers who will also provide helpful information about O*Net, MassCIS, and VIA Character Strengths. Please note, we are trying out a new format to reduce the time we spend on Needs & Leads, so you will have more time for networking. We hope you will be able to join us for this fun and informative program on assessments!


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What is the sum of all numbers $a$ for which the graph of $y=x^2+a$ and the graph of $y=ax$ intersect one time?
for this to be true, x^2+a and ax are equal at only one point. that means that x^2-ax+a's discriminant is 0, which means a^2-4a is 0. that means a is 0 or 4, and the sum is 4.
y = x^2 + a is a parabola
y = ax is a line
If the line intersects the parabola, it will be tangent to the parabola
The slope of a line tangent to the parabola at any point is 2x
So...the slope of the line = a = 2x
So....subbing in for a in both equations, we have that
y = x^2 + 2x
y = 2x (x) = 2x^2
Setting these equal we have that
x^2 + 2x = 2 x^2 rearranging, we have
x^2 - 2x = 0 factor
x( x - 2) = 0 setting each factor to 0 and solving for x produces
x = 0 or x = 2
So....a = 2(0) = 0 or a = 2(2) = 4
And the sum of these values for a is 4


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Aereo, Broadcasters Battle On In Court
Aereo and broadcasters continued their legal wrangling this week over the fate of the service.
There are actually at least two legal venues, a bankruptcy court where Aereo is trying sell its assets or restructure its business, and a district court where broadcasters are suing Aereo for copyright infringement.
There was a flurry of filings as broadcasters, led by Fox, told the court Aereo should not be able to wipe some servers because they could hold evidence in the infringement suit and want the bankruptcy sale stayed until copyright issues are settled.
Broadcasters told the bankruptcy court that it should hold off on setting a schedule for the asset sale or at least modify it to protect broadcasters' interests given that the copyright litigation and bankruptcy cases are "substantially intertwined."
The district court presiding over the copyright case enjoined Aereo from retransmitting broadcasters programming while it decided whether it should make that permanent and extend it to delayed transmissions in addition to live streaming.
Aereo has signaled it might consider restructuring in addition to an asset sale. If Aereo tries to restructure, said the broadcasters in a court filing this week, "the delayed retransmission issue must be addressed to ensure that Aereo's post-confirmation operations do not violate the law and to demonstrate that its plan is feasible."
If it sells the assets to someone else, it should not be able to evade the issue of whether time-delayed retransmissions are lawful in order to attract a buyer willing to gamble on whether the court finds the conduct infringing.
Read the full story at B&C.


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Original Research ARTICLE
Protective Effect of Bioactivity Guided Fractions of Ziziphus jujuba Mill. Root Bark against Hepatic Injury and Chronic Inflammation via Inhibiting Inflammatory Markers and Oxidative Stress
- 1Drug Discovery Laboratory, Institute of Advanced Study in Science and Technology, Guwahati, India
- 2Girijananda Chowdhury Institute of Pharmaceutical Science, Guwahati, India
- 3School of Chinese Medicine, Hong Kong Baptist University, Hong Kong, China
- 4Department of Pathology, Hayat Hospital, Guwahati, India
- 5Jawaharlal Nehru Centre for Advanced Scientific Research, Bengaluru, India
The tribal communities of North Eastern India rely on herbal medicine to cure various disease conditions. Ziziphus jujuba Mill. (Rhamnaceae) is one of such medicinal plants used for curing liver ailments, insomnia, anemia, diarrhea, diabetic complications, cancer, and loss of appetite. The present study was aimed to describe the protective ability of Z. jujuba root bark (ZJRB) against hepatic injury and chronic inflammation. Bioactivity guided fractionation of Z. jujuba methanol extract (ZJME) was performed using different solvents of increasing polarity viz. hexane (ZJHF), chloroform (ZJCF), ethyl acetate (ZJEAF), water (ZJWF), and residue (ZJMR). In vitro antioxidant results revealed that both ZJME and ZJWF possess strong antioxidant activity among all the fractions and mother extract tested. Further, ZJME and ZJWF showed significant protection against CCl4 intoxicated HepG2 cell lines by means of increased cell viability and decreased LDH levels compared to control group. ZJME at 200, 400 mg/kg and ZJWF at 50, 100 mg/kg inhibited the lipid peroxidation and significantly restored the liver function markers (AST, ALT, ALP, LDH, SOD, and CAT) and cytokine levels (TNF-α, Il-1β, and Il-10) in CCl4 induced acute liver damage in rats. All the results were comparable with standard drug silymarin which was further confirmed by histopathology analysis of liver. Similarly, inflammation and increase inflammatory cytokines levels of carrageenan induced paw edema in rats have been refurbished to normal levels on par with the standard drug indomethacin. ZJWF demonstrated potent response than ZJME in all the biological tests conducted. The results of the study signify the ability of ZJRB as good therapeutic agent for liver toxicity and chronic inflammation.
During the metabolism process, numerous number of reacting oxygen species (ROS) like superoxide anions, hydroxy radicals and hydrogen peroxide generates inside the body. This ROS covalently binds to the different cell organelles and involves in causing diseases like cancer, inflammation, arthritis, aging, and cardiac problems (Chen et al., 2011). These reactive intermediates mainly involve in membrane lipid peroxidation which causes genesis of non-alcoholic steatohepatitis and liver carcinoma (Ha et al., 2010). The liver is the complex chemical factory and main organ of the human body which involves in vital functions like cleansing blood, vitamin synthesis, regulation of the supply of body fuel, cholesterol regulation, balancing hormone regulation, and drug metabolism. Different ailments like the virus, chronic alcoholism, and toxic chemicals cause liver damage and failure which is a major problem worldwide (Hou et al., 2013). ROS plays an important role in hepatic fibrogenesis through platelet deriving growth factor. In extreme conditions, ROS activates oxidative stress mediated inflammation and leads to hepatocellular carcinoma (Muriel, 2009).
Inflammation is the defense mechanism by an organism against pathogens, irritants or physical injury, which also implicated in many diseases like multiple sclerosis, rheumatoid arthritis, complications of sepsis, and ischemia-reperfusion injury. All these diseases associated with increased production of ROS due to oxidative stress. Inflammation stimuli trigger the macrophages to produce and release of inflammatory cytokines like tumor necrosis factor alpha (TNF-α) and interleukin 1β (IL-1β) which causes chronic inflammation and pain (Huang et al., 2012; Salzano et al., 2014). There are a few treatment options available in the market for treating the liver ailments and inflammation with undesirable side effects, which demands the new treatment strategies.
Ayurvedic medicine has gained worldwide attention because of its effectiveness without causing unwanted side effects. Presently, the research community is focusing on thoroughgoing pharmacological validation and identification of biomarkers for the treatment of various disease conditions (Feng et al., 2011; Pareek et al., 2013; Choudhury B. et al., 2016). North Eastern India is a part of Indo-Burma biodiversity hotspot known for widespread availability of herbal medicine. Ziziphus jujuba Mill. (Rhamnaceae) is one such plant used by traditional healers for the treatment of liver ailments, cancer, diabetic complications, insomnia, anemia, diarrhea, and loss of appetite (Elaloui et al., 2016). This plant possesses different chemical components like alkaloids, flavonoids, glycosides, and phenolic group of compounds (Yoshikawa et al., 1997; Pawlowska et al., 2000). Z. jujuba was reported for anti-cancer (Huang et al., 2007), anxiolytic (Peng et al., 2000), anti-complementary (Lee et al., 2004), and hypoglycemic (Iganacimuthu and Amalraj, 1998) activities. Different parts of this plant have been using by tribal communities of North East India for the treatment of different disease conditions. Especially root bark of this plant has been using for the treatment of liver toxicity, jaundice, and diabetic complications. These claims were authenticated by Dr. Dinesh Boruah, Senior scientist at North East Indian Ayurvedic Research Institute, Govt. of India, Guwahati, Assam. Since there is no scientific validation of this folklore claim, the present study was aimed to scientifically validate the bioactive guided fractions of Z. jujuba root bark (ZJRB) against liver toxicity and chronic inflammation using in vitro and in vivo methods.
Materials and Methods
Chemical and Drugs
2,2-diphenyl-1-picryl hydrazyl (DPPH), 3-(4,5-dimethyl thiazol-2-yl)-5-diphe-nyl tetrazolium bromide (MTT), trypan blue, trypsin, carrageenan were obtained from Sigma-Aldrich Co, St Louis, MO, USA. Fetal Bovine serum (FBS) and Dulbecco's Modified Eagle medium (DMEM) low glucose were purchased from Invitrogen, Life technologies, USA. Trichloroacetic acid (TCA), thiobarbituric acid (TBA), and ethylenediaminetetraacetic acid (EDTA) were obtained from Hi-Media Lab, Pvt. Ltd, Mumbai. TNF-α, IL-1β, & IL-10 kits were purchased from R&D systems, USA. Diagnostic kits for serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) were purchased from Accurex Biomedical Pvt. Ltd, Mumbai. Superoxide dismutase (SOD) and catalase (CAT) assay kits were procured from Cayman, USA. All the other chemicals used in this study were of analytical grade and obtained from either Sigma-Aldrich or Merck.
Plant Collection and Identification
Ziziphus jujuba plant root bark was collected from Guwahati, Assam (26.1833° N, 91.7333° E) in the month of May 2015 and identified by a taxonomist at North East India Ayurvedic Institute, Guwahati. A voucher specimen number (727/IASST/ 2014) was deposited at herbarium, Drug discovery laboratory, Institute of Advanced Study in Science and Technology (IASST), Guwahati, Assam for future reference.
Extraction and Bioactivity Guided Fractionation
Ziziphus jujuba root bark was shade dried at room temperature (25–27°C) and ground into coarse powder. About 10 kg of the dried powder was subjected to maceration with methanol for 72 h. The Z. jujuba root bark methanol extract (ZJME) was concentrated under pressure using rota evaporator (Buchi, Switzerland) to yield the dry residue. A part of ZJME (400 g) was subjected to gradual fractionation using different solvents of increasing polarity viz, hexane (ZJHF), chloroform (ZJCF), ethyl acetate (ZJEAF), and water (ZJWF). Briefly, 400 g of ZJME was suspended in 1 l of hexane and stirred vigorously by using magnetic stirrer for 24 h at room temperature. Further, the hexane fraction was collected by filtration and the undissolved residue was collected and dried to continue the further fractionation. After extracting with all the solvents, the remaining methanol residue was collected and dried (ZJMR). The schematic representation of bioactivity guided fractionation process is shown in Figure 1. Main mother methanol extract and its five fractions were prepared for testing biological activity and stored at 4°C. All the samples were tested within 3 months of extraction and thawed before use (Kandimalla et al., 2016a).
FIGURE 1. Schematic representation of bioactive guided fractionation of Z. jujuba root bark. ZJME, Z. jujuba root bark methanol extract; ZJHF, Z. jujuba root bark hexane fraction; ZJCF, Z. jujuba root bark chloroform fraction; ZJEAF, Z. jujuba root bark ethyl acetate fraction; ZJWF, Z. jujuba root bark water fraction; ZJMR, Z. jujuba root bark methanol residue.
In vitro Antioxidant Activity
DPPH Radical Scavenging Assay
The radical scavenging activity was evaluated by DPPH assay (Jingsheng et al., 2012). Briefly, 2.7 mL of 0.2 mM DPPH was added to 0.3 mL of the extract solution at various concentrations. The mixture was shaken vigorously and incubated at room temperature for 1 h before the absorbance was measured at 517 nm. The radical scavenging activity was calculated as follows: scavenging rate = [(As - Ai)/As] × 100, where As is the absorbance of pure DPPH and Ai is the absorbance of DPPH in the presence of various extracts. Ascorbic acid at different concentrations identical to the experimental samples was used as references.
Reducing Power Assay
The reducing power of the extracts was estimated using the method by Choudhury B. et al. (2016). Increased concentrations of 0.2 mL extracts were mixed with 2.5 mL of phosphate buffer (0.2 M, pH 6.6) and 2.5 mL of potassium ferricyanide (1%). After incubation at 50°C for 20 min, 2.5 mL of TCA (10%) was added, and each mixture was centrifuged at 1000 rpm for 10 min. Then, 2.5 mL of the supernatant was collected and mixed with 2.5 mL of deionized water and 0.5 mL of ferric chloride (0.1%). The absorbance was measured at 700 nm. The increased absorbance of the reaction mixture indicated increased reducing power. BHT was used as standards for comparison.
Total Antioxidant Activity
Measurement of total antioxidant capacity of plant extracts was done by photochemiluminescence method in the Photochem instrument, Germany. The lipophilic and hydrophilic antioxidants were measured with the commercially available kits from by Photochem (Germany) according to the protocol given by the manufacturer. Total antioxidant activity of plant extracts was expressed in trolox equivalents for lipophilic antioxidants and ascorbic acid equivalents for hydrophilic antioxidants (Kalita H. et al., 2016).
Cell Culture
Human liver hepatoma cells (HepG2) were procured from National Center for Cell Sciences (NCCS), Pune, India. The cells were seeded (1 × 105 cells/T25 flask) and cultured in DMEM with low glucose containing 10% FBS and Penstrap (antibiotic solution) in CO2 Incubator at 37°C. Stock culture was grown in 25 cm2 culture flasks and all experiments were performed in 96 well plates (Life Technologies, USA).
In vitro Cytotoxicity Assay
Tetrazolium salt assay (MTT) was used to determine the cytotoxic concentration of ZJME and ZJWF (Kalita S. et al., 2015, 2016; Kandimalla et al., 2016b; Konwar et al., 2016). Briefly, the cells from the culture flask were trypsinized and seeded in 96-well plate at 5 × 103 cells/well in a culture medium and incubated in the CO2 incubator at 37°C for 24 h. Then the culture medium from the wells was replaced with new media and ZJME and ZJWF at different concentrations (5, 25, 50,125, 250, and 500 μg/mL). After 72 h of incubation in the CO2 incubator at 37°C, media was removed and 20 μl of 4 mg/mL MTT (pH 7.4) was added to each well. The plate was incubated for four more hours and then the supernatant was removed following 100 μl of DMSO was added to each well and further incubated for 30 min to dissolve the formed formazan. Absorbance was read in at 570 nm by using microplate reader (Thermo Scientific, USA). The percentage growth inhibition was calculated by using the following formula-
Protective Effect of Z. jujuba Active Fractions on CCl4 Induced Toxicity in HepG2 Cell Lines
HepG2 cells at 1 × 105 cells/mL were adjusted with DMEM medium containing 10% FBS. To each well of 96 well microtitre plate, 0.1 mL of diluted cell suspension was added and incubated at 37°C for 24 h in the CO2 incubator. After 24 h of incubation, the media was discarded and expose the cells with different drug treatments as follows-
Group-I (Control):
Normal control: Cells were treated with 100 μl of serum free culture medium for 24 h.
DMSO control: Cells were treated with 100 μl of serum free culture medium containing DMSO (0.25% v/v) for 24 h.
Silymarin Control: Cells were treated with 100 μl of serum free culture medium containing silymarin (200 μg/mL) for 24 h.
ZJME Control: Cells were treated with 100 μl of serum free culture medium containing ZJME (200 μg/mL) for 24 h.
ZJWF control: Cells were treated with 100 μl of serum free culture medium containing ZJWF (200 μg/mL) for 24 h.
Group-II (CCl4 treatment): Cells were treated with 100 μl of serum free culture medium containing 1.0% (v/v) CCl4 for 24 h.
Group-III (Standard treatment): Cells were treated with 100 μl of serum free culture medium containing 1.0% (v/v) CCl4 and silymarin at different concentrations (50, 100, or 200 μg/mL) for 24 h.
Group-IV (ZJME treatment): Cells were treated with 100 μl of serum free culture medium containing 1.0% (v/v) CCl4 and ZJME at a concentration of (50, 100, or 200 μg/mL) for 24 h.
Group-V (ZJWF treatment): Cells were treated with 100 μl of serum free culture medium containing 1.0% (v/v) CCl4 and ZJWF at a concentration of (50, 100, or 200 μg/mL) for 24 h.
Cell Viability
Trypan blue exclusion assay (Rambabu and Vijayakumar, 2014) was performed to determine the cell viability. Briefly, after the exposure of cells to different treatments, cells from the wells were trypsinized and centrifuged at 1000 × g for 10 min at 4°C. The pellet was resuspended in 1 mL phosphate buffer saline (PBS), and then 0.1 mL of cell suspension was mixed with 0.1 mL of 0.2% trypan blue. Cells were counted using hemocytometer under light microscope and the percentage of viable cells was determined by using the following formula:
Measurement of Lactate Dehydrogenase (LDH)
After completion of the drug treatment period, culture media from the treatment groups were centrifuged at 2000 rpm for 15 min to separate the supernatant. LDH levels in the supernatant were measured by using Ecoline diagnostic kit as per the instructions are given by the manufacturer. The amount of enzyme catalyzes the conversion of lactate to pyruvate to produce 1.0 μmol of Nicotinamide adenine dinucleotide (NADH) per minute describe as one unit of LDH activity.
Acute Toxicity Studies
Acute oral toxicity studies were performed according to Organization for economic cooperation and development 423 (OECD 423) guidelines to test chemicals. Swiss albino mice of either sex (n = 6) were randomly selected for the study. Animals were kept overnight fasting with free access to water but not food, next day morning single dose of ZJME and ZJWF 2000 mg/kg body weight were administered orally to three animals of each group. Behavioral changes were observed for 24 h after the drug administration and animals were observed for 14 days, if mortality was observed in two out of three animals, then the dose was identified as the toxic dose. If mortality was observed in one animal, experiment was repeated again with the same dose to confirm the toxic dose. On further observation of mortality, the experiment was continued with low doses (300, 50, and 5 mg/kg body weight; Kandimalla et al., 2016d).
Effect of Z. jujuba Active Fraction in CCl4 Intoxicated Rats
Adult male Wistar rats weighing 150–200 g were obtained from the Institute of Advanced Study in Science and Technology (IASST), Guwahati (India). Animals were housed and maintained at 24°C ± 1°C, relative humidity of 45–55% and 12:12 h dark/light cycle. Animals were free to access the water and standard pellet diet (Provimi Animal Nutrition Pvt. Ltd., India) throughout the experiment and all experiments were carried out between 09:00 and 17:00 h. The experimental procedures were approved by the Institutional Animal Ethics Committee (IAEC) of IASST, Guwahati (IASST/IAEC/2014-15/746) and all the experiments conducted in accordance with the CPCSEA (Committee for Purpose of Control and Supervision of Experimentation on Animals) guidelines.
Experimental Design and Drug Treatment
Adult male Wistar rats, a total of 42 animals were randomly divided into seven groups of six animals in each group. All the drug treatment was continued for 14 days and on 14th day single dose of CCl4 1.5 mg/kg, in 1:1 dilution with olive oil was given in i.p. route.
Group I: D.W for 14 days orally + CCl4 1.5 mg/kg on 14th day.
Group II: 0.3% CMC for 14 days orally + CCl4 1.5 mg/kg on 14th day.
Group III: Silymarin 100 mg/kg in 0.3% CMC for 14 days orally + CCl4 1.5 mg/kg on 14th day.
Groups IV and V: ZJME at 200 and 400 mg/kg in 0.3% CMC for 14 days orally + CCl4 1.5 mg/kg on 14th day.
Groups VI and VII: ZJWF at 50 and 100 mg/kg in 0.3% CMC for 14 days orally + CCl4 1.5 mg/kg on 14th day.
After 48 h of CCl4 administration, all the animals were sacrificed to collect blood and liver for biochemical estimation and histopathology analysis.
Serum Biochemical Estimation
Blood was collected by retro orbital route under mild anesthesia and serum was separated by centrifugation at 2000 rpm for 10 min and stored at -80°C for further use. TNF-α, IL-1β, and IL-10 were measured by using ELISA kits from R&D systems as per the instructions are given by the manufacturer, each sample was performed in duplicate and results were expressed in Pg/mL. The levels of hepatobiliary enzymes like AST, ALT, ALP, LDH, and bilirubin were estimated by commercially available assay kits from Accurex, India.
Measurement of Liver Antioxidant Enzymes and Lipid Peroxidation
After blood collection animals from all the groups were sacrificed using an overdose of ether anesthesia and liver was collected immediately and divided into two parts. One part was stored in 10% buffered formaldehyde for histopathology analysis and the second part was frozen in liquid nitrogen for biochemical estimation. The liver homogenate was prepared with 50 mM cold potassium phosphate buffer (pH 7.4) and centrifuged at 3000 rpm for 15 min (Jain et al., 2008). Supernatant was collected for the estimation of SOD, CAT using assay kits from Cayman, USA and TBA reacting substances (TBARS; Fan et al., 2009; Xia et al., 2013). All the experiments were conducted at 4°C.
Histopathology Examination of Liver
Livers were stored in 10% buffered formaldehyde and preserved for at least 24 h, dehydrated gradually with ethanol (70–100%), cleared in xylene and embedded in paraffin. Sections of 5 μm were prepared and stained with hematoxylin and eosin to examine under the microscope (10×) for histopathological changes (Bhardwaj et al., 2016; Choudhury A.J. et al., 2016).
Effect of Z. jujuba Fractions against Carrageenan Induced Paw Edema
Adult male Wistar rats (N = 36), weighing 200–220 g were randomly divided into six groups (n = 6). Inflammation was induced in overnight fasted animals by intraplantar injection of 1% carrageenan (0.05 mL) to right hind paw (Arumugam et al., 2008). The following drug treatments were given to the animals before injecting the carrageenan.
Group I: Control animals (0.3% CMC orally)
Group II: Treatment with 10 mg/kg indomethacin orally.
Groups III and IV: Treatment with ZJME 200 mg/kg and 400 mg/kg orally.
Groups V and VI: Treatment with ZJWF 50 mg/kg and 100 mg/kg orally.
Water plethysmometer (Harvard apparatus, Panlab, Spain) was used to measure the paw volume at different time intervals (0, 1, 3, and 5 h) after the induction of inflammation. At the end of the experiment, blood was collected through retro orbital route from all the animals under mild anesthesia to measure the serum inflammatory markers. All the animals were sacrificed at the end of the experiment.
Measurement of Inflammatory Cytokine Levels
Serum was separated from blood by centrifugation at 2000 rpm for 10 min. Inflammatory cytokines like TNF-α and Il-1β were measured using ELISA kits from R&D systems, the USA as per the protocol is given by the manufacturer. All the treatments were done at 4°C and results were presented in pg/mL (Kandimalla et al., 2016c).
Statistical Analysis
All the results were expressed as mean ± SD. One way ANOVA followed by Tukey's multiple comparison tests were used to compare the different parameters between the groups. A p-value < 0.05 was considered as significant.
Phytochemical Investigation of ZJME and ZJWF
Both ZJME and ZJWF were screened for phytochemicals to determine the nature of the chemical constituents present using following methods-
Test for alkaloids: 0.5 g of the test sample was stirred with 5 ml of aqueous HCl acid (1%) on the water bath at 95°C. The reaction mixture was filtered using Whatman filter paper and few drops of dragendorff's reagent were added to 1 ml of the filtrate. Formation of orange red precipitate was considered for the presence of alkaloids (Adegoke et al., 2010).
Test for flavonoids: To 0.5 g of the test sample 5 ml of ethyl acetate was added and heated in a water bath and the reaction mixture was filtered. To 1 ml of the filtrate, few drops of ammonia solution were added and shaken vigorously. Formation of yellow color indicates the presence of flavonoids (Kalita et al., 2011).
Test for phenolic compounds: About 50 mg of the test sample was dissolved in 5 ml of distilled water. To this mixture, 5% ferric chloride solution was added. Formation of dark green color indicated the presence of phenolic compounds (Kalita et al., 2011).
Test for saponins: To a pinch of test sample 2–3 ml of distilled water was added and shake vigorously. Formation of foam indicated the presence of saponins (Adegoke et al., 2010; Kalita et al., 2011).
Test for steroids: To 0.5 g of test sample 2 ml of acetic anhydride and 2 ml concentrated sulfuric acid was added. Change of color from violet to blue indicates the presence of steroids (Boxi et al., 2010; Kalita et al., 2011).
Test for terpenes: To 0.5 g of test sample 3 ml of concentrated sulfuric acid and 2 ml of chloroform was added. Formation of reddish brown ring confirms the presence of terpenes (Obianime and Uche, 2008).
Test for cardiac glycosides: Keller–Killani test was performed to determine the presence of cardiac glycosides. To 0.5 g of test sample, 2 ml of glacial acetic acid and one drop of ferric chloride was added. The reaction mixture was underlaid with 1 ml of concentrated sulphuric acid. Formation of the brown ring at the interface indicates the presence of cardiac glycosides.
Test for oils and fats: A small amount of test sample was pressed between the two filter papers. Formation of oil stain indicates the presence of fats and oils (Kalita et al., 2011).
Test for proteins and amino acids: 50 mg of the test sample was mixed with few ml of diluted HCl and filtered. To the filtrate, few drops of ninhydrin solution (10 mg of ninhydrin in 200 ml of Acetone) was added. Formation of purple color indicates the presence of proteins and amino acids.
Total Phenolic and Flavonoid Content of ZJME and ZJWF
The total phenolic content of the ZJME and ZJWF were determined using the Folin–Ciocalteu assay. The results are expressed as grams of gallic acid equivalents per 100 g of dry extract. The total flavonoid content was determined by a colorimetric assay using rutin as a standard. The results are expressed as grams of rutin equivalents per 100 g of dry extract (Kandimalla et al., 2016d).
Results and Discussion
Extraction and Fractionation
Maceration of ZJRB (10 kg) with methanol yields 524 g of ZJME. Further, fractionation of ZJME (400 g) with different solvents of increasing polarity yields as follows- ZJHF (38 g), ZJCF (74 g), ZJEAF (27 g), and ZJWF (124 g). After the successful fractionation with all the solvents, the left out residue was also collected ZJMR (116 g). During the fractionation process, some amount of ZJME was lost (21 g).
In vitro Antioxidant Activity of Z. jujuba Fractions
DPPH Radical Scavenging Activity
2,2-diphenyl-1-picryl hydrazyl is a free radical; the ability of test substance to neutralize the DPPH radical is considered as its anti-oxidant ability which is a widely accepted method to determine the in vitro antioxidant capacity (Liu et al., 2013). The free radical scavenging ability of Z. jujuba methanol extract (ZJME) and its fractions was given in Figure 2. Among all the fractions tested Z. jujuba water fraction (ZJWF) and ZJME showed potent response, which is comparable with standard ascorbic acid. At higher concentration, ZJWF showed potent activity than standard ascorbic acid. Z. jujuba hexane fraction (ZJHF), Z. jujuba chloroform fraction (ZJCF), Z. jujuba ethyl acetate fraction (ZJEAF) and Z. jujuba methanol residue (ZJMR) also exhibited dose dependent antioxidant activity but not up to the activity of ZJWF and ZJME. This result of this experiment suggests that both ZJWF and ZJME have the strong antioxidant ability.
FIGURE 2. DPPH radical scavenging activity of different fractions of Z. jujuba root bark. ZJME, Z. jujuba root bark methanol extract; ZJHF, Z. jujuba root bark hexane fraction; ZJCF, Z. jujuba root bark chloroform fraction; ZJEAF, Z. jujuba root bark ethyl acetate fraction; ZJWF, Z. jujuba root bark water fraction; ZJMR, Z. jujuba root bark methanol residue; Vc, Ascorbic acid. All the results were expressed in mean ± SD (n = 3).
Reducing Power Assay
The ability of a test substance that reduce the Fe3+ to Fe2+ is considered as reducing power ability which is directly related to its antioxidant capacity. Reducing power ability of the substance can be measured spectrophotometrically (Meir et al., 1995). ZJME and ZJWF showed strong antioxidant activity and the later one is potent among all the fractions and extract which is comparable with standard BHT (Figure 3). So we selected ZJME and ZJWF for further pharmacological analysis.
FIGURE 3. Reducing power ability of different fractions of Z. jujuba root bark. ZJME, Z. jujuba root bark methanol extract; ZJHF, Z. jujuba root bark hexane fraction; ZJCF, Z. jujuba root bark chloroform fraction; ZJEAF, Z. jujuba root bark ethyl acetate fraction; ZJWF, Z. jujuba root bark water fraction; ZJMR, Z. jujuba root bark methanol residue; BHT, Butylated hydroxyl toluene. All the results were expressed in mean ± SD (n = 3).
Total Antioxidant Activity
Total antioxidant capacity of a test substance was measured by photochemiluminescence method in the Photochem instrument. This reaction depends on antioxidant concentration which follows the Guldberg–Waage law. The reaction of radicals with antioxidant molecules forms a stable compound which can be measured spectrophotometrically (Hic and Balic, 2012). Total antioxidant activity of ZJME and ZJWF was 69.76 nmol trolax equivalents and 94.27 nmol ascorbic acid equivalents, respectively.
Cytotoxicity Effect of ZJME and ZJWF in HepG2 Cells
ZJME and ZJWF at 5, 25, 50,125, and 250 μg/ml did not show the significant effect on HepG2 cell growth but at the highest dose 500 μg/mL both exhibited growth inhibition. So, we selected 50, 100, and 200 μg/ml of ZJME and ZJWF for evaluation of protective effect against CCl4 intoxicated HepG2 cells.
Protective Effect of ZJME and ZJWF in CCl4 Induced Toxicity on HepG2 Cells
In this study, the hepatoprotective ability of ZJME and ZJWF have ascertained in CCl4 intoxicated HepG2 cell lines. In this type of studies, HepG2 cells have been used as a substitute to hepatocytes. CCl4 is a well know toxin that induces lipid peroxidation in HepG2 cells and cause severe damage, which regarded as a convenient method to screen the medicinal plants extract and isolates against hepatotoxicity (Krithika et al., 2013). Table 1. shows the results of cell viability and LDH levels of HepG2 cells. Treatment with CCl4 significantly reduced the HepG2 cell viability and increased the LDH leakage into the culture media. ZJME and ZJWF treatment along with CCl4 reduce the toxic effect of CCl4 which was confirmed by increased cell viability and decreased LDH leakage. Results of this experiment demonstrated the hepatoprotective ability of ZJME and ZJWF against CCl4 intoxication in HepG2 cells; this might be due to the membrane stabilization. ZJWF showed potent response than ZJME, which explains that polar chemical components of ZJRB responsible for hepatoprotection. For further confirmation, we performed in vivo experiments.
Acute Toxicity Studies
After single large dose (2000 mg/kg) administration of ZJME and ZJWF, we observed the animals for toxic effects and behavioral changes. Both ZJME and ZJWF did not alter the heart rate, body temperature, respiratory rate, locomotor activity, salivation, corneal reflex, grip strength, abdominal tone, body tone, piloerection, tail elevation, tremors, convulsions, and twitches in animals. After 14 days of the observation period, no mortality was observed in the animals. The results of this study explain that both ZJME and ZJWF non-toxic to animals and we further continued for activity evaluation.
Effect of ZJME and ZJWF against CCl4 Induced Hepatic Damage
Effect on Serum Hepatobiliary Enzyme Levels
Table 2 shows the effect of ZJME and ZJWF on serum hepatobiliary enzymes like AST, ALT, ALP, and LDH levels. CCl4 treatment significantly raised all these serum biochemical markers. The 14 days of pretreatment with ZJME, ZJWF, and standard drug silymarin protects the liver from CCl4, which clearly observed by restored levels of all these markers. CCl4 is a well-known hepatotoxin that damages the plasma membrane of liver cells and causes necrosis. In deep inside the living system CCl4 converts into trichloromethyl radical (CCl3.), an active metabolite which covalently bonds with sulfhydryl groups in protein thiols of hepatocytes and causes lipid peroxidation and necrosis. During this necrotic stage, hepatobiliary enzymes release into blood circulation which is the reason behind the high enzyme levels after CCl4 treatment (Vuda et al., 2012). Results of this study demonstrated the hepatoprotective ability of both ZJME and ZJWF, where the latter one is more potent than former which states that polar chemical components of Z. jujuba were responsible for this activity.
Effect on Lipid Peroxidation and Liver Antioxidant Enzymes
Superoxide dismutase and catalase are two powerful antioxidant enzymes that produce inside the body. SOD decomposes the superoxide radical (O2⋅-) into hydrogen peroxide (H2O2), which subsequently removed to water by CAT in the peroxisomes. CCl4 induction causes the significant decrease in antioxidant enzymes like SOD and CAT which leads to the lipid peroxidation and toxicity (Cheng et al., 2013). Treatment with CCl4 caused significantly (P < 0.001) increased the lipid peroxidation and declined SOD and CAT enzymes levels (Table 2). Treatment with ZJME, ZJWF, and standard drug silymarin showed significant protection from CCl4 which was demonstrated by reduced lipid peroxidation and high levels of antioxidant enzymes. ZJWF showed potent activity than ZJME which refers that the polar chemical components of ZJRB were responsible for this activity.
Effect on Serum Cytokine Levels
CCl4 metabolism triggers the liver kupffer cells, which leads to activation of inflammatory cytokines like TNF-α and IL-1β and reduces the anti-inflammatory cytokine IL-10. IL-1β is a powerful inflammatory cytokine, stimulates the macrophages and involves in prostaglandins production and neutrophil infiltration. Activation of this inflammatory cytokine cascade cause significant damage to the hepatocytes (Edwards et al., 1993; Ishiyama et al., 1995). CCl4 injection significantly raised the TNF-α, Il-1β, and reduced the IL-10 levels in the rats. ZJME and ZJWF treatment dose dependently reduced the TNF-α (Figure 4), IL-1β (Figure 5) production and increased the IL-10 levels (Figure 6) in animals intoxicated with CCl4. ZJWF was found to be more potent than ZJME which indicated that the active components are polar in nature.
FIGURE 4. Effect of different drug treatment on serum TNF-α levels. All the results were expressed in mean ± SD (n = 6). $$$P < 0.001 in comparison of CCl4 alone treated animals with normal animals. ∗∗∗P < 0.001 and ∗∗P < 0.01 in comparison of drug treated animals with CCl4 alone treated animals. ZJME, Z. jujuba root bark methanol extract; ZJWF, Z. jujuba root bark water fraction.
FIGURE 5. Effect of different drug treatment on serum IL-1β levels. All the results were expressed in mean ± SD (n = 6). $$$P < 0.001 in comparison of CCl4 alone treated animals with normal animals. ∗∗∗P < 0.001 in comparison of drug treated animals with CCl4 alone treated animals. ZJME, Z. jujuba root bark methanol extract; ZJWF, Z. jujuba root bark water fraction.
FIGURE 6. Effect of different drug treatment on serum IL-10 levels. All the results were expressed in mean ± SD (n = 6). $$$P < 0.001 in comparison of CCl4 alone treated animals with normal animals. ∗∗∗P < 0.001, ∗∗P < 0.01, and ∗P < 0.05 in comparison of drug treated animals with CCl4 alone treated animals. ZJME, Z. jujuba root bark methanol extract; ZJWF, Z. jujuba root bark water fraction.
Effect on Histopathology of Rat Liver
In support of the hepatoprotective response of ZJME and ZJWF, liver tissues from different treatment groups undergone for histopathology analysis. Hepatocytes with normal texture were observed with no cell infiltrate in control animals (Figure 7A). CCl4 intoxication causes demolishment of hepatocytes which was evidenced by the formation of bridging necrosis, collagen accumulation, large septa, and chronic inflammation (Figure 7B). Standard drug silymarin, ZJME (400 mg/kg) and ZJWF (100 mg/kg) pretreatment to the animals showed significant protection from CCl4, where normal liver architecture was observed with no necrosis and mild inflammation (Figures 7C–E). This histopathology data confirms the hepatoprotective ability of ZJRB and its polar chemical components were responsible for the activity.
FIGURE 7. Effect of drug treatment on liver histopathology of CCl4 intoxicated rats. (A) Liver of control rat with no CCl4 treatment showing normal hepatocytes with no inflammatory cell infiltrate; ×10. (B) Liver of rat treated with CCl4 showing formation of septa and bridging necrosis with chronic inflammatory cells; ×10. (C) Liver of rat treated with CCl4 and 100 mg/kg silymarin showing normal hepatocytes with no inflammation and necrosis; ×10. (D) Liver of rat treated with CCl4 and 400 mg/kg of ZJME showing mild periportal inflammation and no necrosis; ×10. (E) Liver of rat treated with CCl4 and 100 mg/kg ZJWF showing absence of inflammatory cells or necrosis; ×10. ZJME, Z. jujuba root bark methanol extract; ZJWF, Z. jujuba root bark water fraction.
Effect of ZJME and ZJWF on Carrageenan Induced Paw Edema
Effect on Paw Volume
Inflammation occurs in two step process, in first step histamine and serotonin releases whereas the second step involves the secretion of prostaglandins and lysosomal bodies, which are the primary target for anti-inflammatory drugs in the market. Carrageenan induced inflammation in experimental animals is the widely accepted method to test the orally injected anti-inflammatory drugs (Marroquin-Segura et al., 2009). ZJME, ZJWF, and standard drug indomethacin reduced the paw volume significantly after carrageenan injection compared to non-treated animals (Figure 8). ZJWF showed potent response than ZJME which clearly indicates that polar chemical components of the ZJRB are responsible for this pharmacological response.
FIGURE 8. Effect of Z. jujuba root bark fractions on carrageenan induced paw edema over 5 h. All the results were expressed in mean ± SD (n = 6). S is p < 0.05 in comparison of drug treated animals with saline treated animals. ZJME, Z. jujuba root bark methanol extract; ZJWF, Z. jujuba root bark water fraction.
Effect on Serum Cytokine Levels
IL-1β and TNF-α are two powerful inflammatory cytokines which play a major role in the activation of the prostaglandin and macrophages. Intraplantar injection of carrageenan to rats causes the significant rise in TNF-α and IL-1β levels. Pretreatment of indomethacin, ZJME, and ZJWF before carrageenan shot inhibits the cytokines levels (Figures 9 and 10) compared to non-treated animals. ZJWF showed potent response than ZJME which demonstrates that anti-inflammatory compounds of ZJRB were polar in nature.
FIGURE 9. Effect of different drug treatment on serum TNF-α levels of carrageenan induced paw edema rats. All the results were expressed in mean ± SD (n = 6). ∗∗∗P < 0.001 and ∗∗P < 0.01 in comparison of drug treated animals with saline treated animals. ZJME, Z. jujuba root bark methanol extract; ZJWF, Z. jujuba root bark water fraction.
FIGURE 10. Effect of different drug treatment on serum IL-1β levels of carrageenan induced paw edema rats. All the results were expressed in mean ± SD (n = 6). ∗∗∗P < 0.001 and ∗P < 0.05 in comparison of drug treated animals with saline treated animals. ZJME, Z. jujuba root bark methanol extract; ZJWF, Z. jujuba root bark water fraction.
Phytochemical Investigation of ZJME and ZJWF
The pharmacologically active fractions ZJME and ZJWF were subjected to qualitative phytochemical analysis to determine the nature of the active compounds. ZJME showed the presence of alkaloids, flavonoids, phenolic compounds, terpenes, saponins, and steroids where alkaloids and steroids were absent in ZJWF (Table 3). Based on these findings further investigation of bioactive compounds isolation is in progress in our laboratory and will be reported in our further communications.
Total Phenolic and Flavonoid Content of ZJME and ZJWF
Antioxidant capacity of a plant fraction or extract correlates with the presence of the phenolic type of compounds (Bravo, 1998; Huang et al., 2010; Chen et al., 2011). Total phenolic content of ZJME and ZJWF was found to be 246.4 ± 4.37 mg and 322.6 ± 3.72 mg gallic acid equivalents/g extract, respectively. The total flavonoid content of ZJME and ZJWF was found to be 4.9 ± 0.42 mg and 6.2 ± 0.46 mg rutin equivalents/g extract, respectively. Results of this study explain that ZJWF contains rich amount of phenolic and flavonoid compounds than ZJME. Further investigation on enrichment and isolation of bioactive compounds is ongoing in our laboratory.
This study demonstrated the protective ability of ZJRB against hepatotoxicity and inflammation. Hepatoprotective activity may be attributing to its strong antioxidant ability which reduced the oxidative stress and lipid peroxidation. Further, ZJRB showed anti-inflammatory activity through inhibiting the pro-inflammatory cytokines. Water fraction of methanol extract showed potent response toward all the in vitro and in vivo tests conducted, this may be due to polar chemical components of the plant. Therefore the study scientifically validates the traditional usage of ZJRB for the treatment of liver ailments and inflammation. Successful isolation of the biomarker can potentially contribute toward the development of new drug entity.
Author Contributions
RK designed the whole study and performed all the in vitro and in vivo experiments. SK, BC, SM, and BK helped RK in different phases of the study. SD contributed towards study design, supervision and compiling the results. KK performed and analyzed the histopathology of liver tissue. RD helped in literature survey and study design. JK contributed towards the study design, manuscript correction, results compiling, and supervision of the whole study.
Conflict of Interest Statement
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
The results of control, toxin, and standard treated groups have been used in another study of ours (DOI: 10.3389/fphar.2016.00168).
The authors are thankful to Department of Science and Technology (DST), New Delhi, India for financial assistance. RK would like to thank Prof. N. C. Talukdar, Director, Institute of Advanced Study in Science and technology for his motivation and support.
Adegoke, A. A., Iberi, P. A., Akinpelu, D. A., Aiyegoro, O. A., and Mboto, C. I. (2010). Studies on phytochemical screening and antimicrobial potentials of Phyllanthus amarus against multiple antibiotic resistant bacteria. Int. J. Appl. Res. Nat. Prod. 3, 6–12.
Arumugam, P., Priya, N. G., Subathra, M., and Ramesh, A. (2008). Anti-inflammatory activity of four solvent fractions of ethanol extract of Mentha spicata L. investigated on acute and chronic inflammation induced rats. Environ. Toxicol. Pharmacol. 26, 92–95. doi: 10.1016/j.etap.2008.02.008
Bhardwaj, N., Singh, Y. P., Devi, D., Kandimalla, R., Kotoky, J., and Mandal, B. B. (2016). Potential of silk fibroin/chondrocyte constructs of muga silkworm Antheraea assamensis for cartilage tissue engineering. J. Mater. Chem. B 4, 3670–3684. doi: 10.1039/C6TB00717A
Boxi, M., Rajesh, Y., Raja Kumar, V., Praveen, B., and Mangamma, K. (2010). Extraction, phytochemical screening and in-vitro evaluation of anti-oxidant properties of Commicarpus chinesis (aqueous leaf extract). Int. J. Pharm. Biosci. 1, 537–547.
Bravo, L. (1998). Polyphenols: chemistry, dietary sources, metabolism, and nutritional significance. Nutr. Rev. 56, 317–333. doi: 10.1111/j.1753-4887.1998.tb01670.x
Chen, Y., Huang, B., He, J., Han, L., Zhan, Y., and Wang, Y. (2011). In vitro and in vivo antioxidant effects of the ethanolic extract of Swertia chirayita. J. Ethnopharmacol. 136, 309–315. doi: 10.1016/j.jep.2011.04.058
Cheng, N., Ren, N., Gao, H., Lei, X., Zheng, J., and Cao, W. (2013). Antioxidant and hepatoprotective effects of Schisandra chinensis pollen extract on CCl4-induced acute liver damage in mice. Food Chem. Toxicol. 55, 234–240. doi: 10.1016/j.fct.2012.11.022
Choudhury, A. J., Gogoi, D., Chutia, J., Kandimalla, R., Kalita, S., Kotoky, J., et al. (2016). Controlled antibiotic-releasing Antheraea assama silk fibroin suture for infection prevention and fast wound healing. Surgery. 159, 539–547. doi: 10.1016/j.surg.2015.07.022
Choudhury, B., Kandimalla, R., Monisha, J., Bharali, R., Kunnumakkara, A. B., Kalita, K., et al. (2016). Anticancer activity of Garcinia morella on T-cell murine lymphoma via apoptotic induction. Front. Pharmacol. 7:3. doi: 10.3389/fphar.2016.00003
Edwards, M. J., Keller, B. J., Kauffman, F. C., and Thurman, R. G. (1993). The involvement of Kupffer cells in carbon tetrachloride toxicity. Toxicol. Appl. Pharm. 119, 275–279. doi: 10.1006/taap.1993.1069
Elaloui, M., Laamouri, A., Ennajah, A., Cerny, M., Mathieu, C., Vilarem, G., et al. (2016). Phytoconstituents of leaf extracts of Ziziphus jujuba Mill. plants harvested in Tunisia. Ind. Crop Prod. 83, 133–139. doi: 10.1016/j.indcrop.2015.11.029
Fan, G., Tang, J. J., Bhadauria, M., Nirala, S. K., Zhou, B., Li, Y., et al. (2009). Resveratrol ameliorates carbon tetrachloride-induced acute liver injury in mice. Environ. Toxicol. Pharmacol. 28, 350–356. doi: 10.1016/j.etap.2009.05.013
Feng, Y., Wang, N., Ye, X., Li, H., Feng, Y., Cheung, F., et al. (2011). Hepatoprotective effect and its possible mechanism of Coptidis rhizome aqueous extract on carbon tetrachloride-induced chronic liver hepatotoxicity in rats. J. Ethnopharmacol. 138, 683–690. doi: 10.1016/j.jep.2011.09.032
Ha, H. L., Shin, H. J., Feitelson, M. A., and Yu, D. Y. (2010). Oxidative stress and antioxidants in hepatic pathogenesis. World J. Gastroenterol. 28, 6035–6043. doi: 10.3748/wjg.v16.i48.6035
Hic, P., and Balic, J. (2012). Effect of sample dilution on estimated values of antioxidant capacity by photochemiluminiscence method. Acta Univ. Agric. Silvic. Mendel. Brun. 8, 67–72. doi: 10.11118/actaun201260080067
Hou, F., Zhang, R., Zhang, M., Su, D., Wei, Z., Deng, Y., et al. (2013). Hepatoprotective and antioxidant activity of anthocyanins in black rice bran on carbon tetrachloride-induced liver injury in mice. J. Funct. Foods 5, 1705–1713. doi: 10.1016/j.jff.2013.07.015
Huang, B., Ban, X., He, J. S., Tong, J., Tian, J., and Wang, Y. (2010). Hepatoprotective and antioxidant activity of ethanolic extracts of edible lotus (Nelumbo nucifera Gaertn.) leaves. Food Chem. 120, 873–878. doi: 10.1016/j.foodchem.2009.11.020
Huang, G., Pan, C., Liu, F., Wu, T., and Wu, C. (2012). Anti-inflammatory effects of ethanolic extract of Antrodia salmonea in the lipopolysaccharide-stimulated RAW246.7 macrophages and the k-carrageenan-induced paw edema model. Food Chem. Toxicol. 50, 1485–1493. doi: 10.1016/j.fct.2012.01.041
Huang, X., Kojima-Yuasa, A., Norikura, T., Kennedy, D. O., Hasuma, T., and Matsui-Yuasa, I. (2007). Mechanism of the anti-cancer activity of Zizyphus jujuba in HepG2 cells. Am. J. Chin. Med. 35, 517–523. doi: 10.1142/S0192415X0700503X
Iganacimuthu, S., and Amalraj, T. (1998). Effect of leaf extract of Zizyphus jujuba on diabetic rats. Indian J. Pharmacol. 30, 107–112.
Ishiyama, H., Sato, M., Matsumura, K., Sento, M., Keiki, O., and Hobara, T. (1995). Proliferation of hepatocytes and attenuation from carbon tetrachloride hepatotoxicity by gadolinium chloride in rats. Basic Clin. Pharmacol. Toxicol. 77, 293–298. doi: 10.1111/j.1600-0773.1995.tb01030.x
Jain, A., Soni, M., Deb, L., Jain, A., Rout, S. P., Gupta, V. B., et al. (2008). Antioxidant and hepatoprotective activity of ethanolic and aqueous extracts of Momordica dioica Roxb. leaves. J. Ethanopharmacol. 115, 61–66. doi: 10.1016/j.jep.2007.09.009
Jingsheng, H., Huang, B., Xiaoquan, B., Jun, T., Ling, Z., and Youwei, W. (2012). In vitro and in vivo antioxidant activity of the ethanolic extract from Meconopsis quintuplinervia. J. Ethnopharmacol. 141, 104–110. doi: 10.1016/j.jep.2012.02.006
Kalita, H., Boruah, D. C., Deori, M., Hazarika, A., Sarma, R., Kumari, S., et al. (2016). Antidiabetic and antilipidemic effect of Musa balbisiana root extract: a potent agent for glucose homeostasis in Streptozotocin induced diabetic rat. Front. Pharmacol. 7:102. doi: 10.3389/fphar.2016.00102
Kalita, S., Devi, B., Kandimalla, R., Sharma, K. K., Sharma, A., Kalita, K., et al. (2015). Chloramphenicol encapsulated in poly-ε- caprolactone–pluronic composite: nanoparticles for treatment of MRSA-infected burn wounds. Int. J. Nanomed. 10, 2971–2984. doi: 10.2147/IJN.S75023
Kalita, S., Kandimalla, R., Sharma, K. K., Kataki, A. C., Deka, M., and Kotoky, J. (2016). Amoxicillin functionalized gold nanoparticles reverts MRSA resistance. Mater. Sci. Eng. C 61, 720–727. doi: 10.1016/j.msec.2015.12.078
Kalita, S., Kumar, G., Karthik, L., and Rao, K. V. B. (2011). Phytochemical composition and in vitro hemolytic activity of Lantana camara L. (Verbenaceae) leaves. Pharmacologyonline 1, 59–67.
Kandimalla, R., Dash, S., Kalita, S., Choudhury, B., Malampati, S., Kalita, K., et al. (2016a). Bioactive guided fractions of Annona reticulata L. bark: protection against liver toxicity and inflammation through inhibiting oxidative stress and proinflammatory cytokines. Front. Pharmacol. 7:168. doi: 10.3389/fphar.2016.00168
Kandimalla, R., Kalita, S., Choudhury, B., Devi, D., Kalita, D., Kalita, K., et al. (2016b). Fiber from ramie plant (Boehmeria nivea): a novel suture biomaterial. Mater. Sci. Eng. C 62, 816–822. doi: 10.1016/j.msec.2016.02.040
Kandimalla, R., Kalita, S., Choudhury, B., Kalita, K., Dash, S., and Kotoky, J. (2016c). Chemical composition and anti-candidiasis mediated wound healing property of Cymbopogon nardus essential oil on chronic diabetic wounds. Front. Pharmacol. 7:198. doi: 10.3389/fphar.2016.00198
Kandimalla, R., Kalita, S., Saikia, B., Choudhury, B., Singh, Y. P., Kalita, K., et al. (2016d). Antioxidant and hepatoprotective potentiality of Randia dumetorum Lam. leaf and bark via inhibition of oxidative stress and inflammatory cytokines. Front. Pharmacol. 7:205. doi: 10.3389/fphar.2016.00205
Konwar, A., Kalita, S., Kotoky, J., and Chowdhury, D. (2016). Chitosan-iron oxide coated graphene oxide nanocomposite hydrogel: a robust and soft antimicrobial bio-film. ACS Appl. Mater. Interfaces 8, 20625–20634. doi: 10.1021/acsami.6b07510
Krithika, R., Verma, R. J., and Shrivastav, P. S. (2013). Antioxidative and cytoprotective effects of andrographolide against CCl4-induced hepatotoxicity in HepG2 cells. Hum. Exp. Toxicol. 32, 530–543. doi: 10.1177/0960327112459530
Lee, S. M., Park, J. G., Lee, Y. H., Lee, C. G., Min, B. S., Kim, J. H., et al. (2004). Anti-complementary activity of triterpenoides from fruits of Zizyphus jujuba. Biol. Pharm. Bull. 27, 1883–1892. doi: 10.1248/bpb.27.1883
Liu, J., Jia, L., Kan, J., and Jin, C. (2013). In vitro and in vivo antioxidant activity of ethanolic extract of white button mushroom (Agaricus bisporus). Food Chem. Toxicol. 51, 310–316. doi: 10.1016/j.fct.2012.10.014
Marroquin-Segura, R., Flores-Pimentel, M., Carreón-Sánchez, R., Garcia-Burciaga, M. M., Mora-Guevara, J. L. A., Aguilar-Contreras, A., et al. (2009). The effect of the aqueous extract of Helietta parvifolia A. Gray (Rutaceae) stem bark on carrageenan-induced paw oedema and granuloma tissue formation in mice. J. Ethnopharmacol. 124, 639–641. doi: 10.1016/j.jep.2009.06.004
Meir, S., Kanner, J., Akiri, B., and Hadas, S. P. (1995). Determination and involvement of aqueous reducing compounds in oxidative defense systems of various senescing leaves. J. Agric. Food Chem. 43, 1813–1817. doi: 10.1021/jf00055a012
Muriel, M. (2009). Role of free radicals in liver diseases. Hepatol. Int. 3, 526–536. doi: 10.1007/s12072-009-9158-6
Obianime, A. W., and Uche, F. I. (2008). The phytochemical screening and effects of methanolic extract of Phyllanthus amarus leaf on the biochemical parameters of male guinea pigs. J. Appl. Sci. Environ. Manage. 12, 73–77.
Pareek, A., Ashok, G., Roshan, I., and Nagori, B. P. (2013). Antioxidant and hepatoprotective activity of Fagonia schweinfurthii (Hadidi) Hadidi extract in carbon tetrachloride induced hepatotoxicity in HepG2cell line and rats. J. Ethnopharmacol. 150, 973–981. doi: 10.1016/j.jep.2013.09.048
Pawlowska, A. M., Camangi, F., Bader, A., and Braca, A. (2000). Flavonoids of Zizyphus jujuba and Zizyphus spina-christi (L) wild (Rhamnaceae) fruits. Food Chem. 112, 858–862. doi: 10.1016/j.foodchem.2008.06.053
Peng, W. H., Hsieh, M. T., Lee, Y. S., Lin, Y. C., and Liao, J. (2000). Anxiolytic effect of seed of Ziziphus jujuba in mouse models of anxiety. J. Ethnopharmacol. 72, 435–439. doi: 10.1016/S0378-8741(00)00255-5
Rambabu, V., and Vijayakumar, S. (2014). In vitro cytotoxicity perspective of diazepinomicin (ECO-4601) on human hepatoma cell line (HepG2). Biomed. Aging Pathol. 4, 65–70. doi: 10.1016/j.biomag.2013.10.004
Salzano, S., Checconi, P., Hanschmann, E., Lillig, C. H., Bowler, L. D., Chan, P., et al. (2014). Linkage of inflammation and oxidative stress via release of glutathionylated peroxiredoxin-2, which acts as a danger signal. Proc. Natl. Acad. Sci. U.S.A. 111, 12157–12162. doi: 10.1073/pnas.1401712111
Vuda, M., D'Souza, R., Upadhya, S., Kumar, V., Rao, N., Kumar, V., et al. (2012). Hepatoprotective and antioxidant activity of aqueous extract of Hybanthus enneaspermus against CCl4-induced liver injury in rats. Exp. Toxicol. Pathol. 64, 855–859. doi: 10.1016/j.etp.2011.03.006
Xia, D. Z., Zhang, P. H., Fu, Y., Yu, W. F., and Ju, M. T. (2013). Hepatoprotective activity of puerarin against carbon tetrachloride-induced injuries in rats: a randomized controlled trial. Food Chem. Toxicol. 59, 90–95. doi: 10.1016/j.fct.2013.05.055
Yoshikawa, M., Murakami, T., Ikebata, A., Wakao, S., Murakami, N., Matsuda, H. J. Y., et al. (1997). Bioactive saponins and glycosides. X. On the constituents of Zizyphi spinosi semen, the seeds of Ziziphus jujuba Mill. var. spinosa Hu (1): structures and histamine release-inhibitory effect of jujubosides A1 and C and acetyljujuboside B. Chem. Pharm. Bull. (Tokyo) 45, 1186–1192.
Keywords: liver toxicity, antioxidant, cytokines, inflammation, oxidative stress
Citation: Kandimalla R, Dash S, Kalita S, Choudhury B, Malampati S, Kalita K, Kalita B, Devi R, Kotoky J (2016) Protective Effect of Bioactivity Guided Fractions of Ziziphus jujuba Mill. Root Bark against Hepatic Injury and Chronic Inflammation via Inhibiting Inflammatory Markers and Oxidative Stress. Front. Pharmacol. 7:298. doi: 10.3389/fphar.2016.00298
Received: 29 April 2016; Accepted: 23 August 2016;
Published: 07 September 2016.
Edited by:Adolfo Andrade-Cetto, National Autonomous University of Mexico, Mexico
Reviewed by:Keliang Xie, Tianjin Medical University, China
Pinarosa Avato, University of Bari, Italy
Justin Antony, Jagadguru Sri Shivaratreeshwara College of Pharmacy, India
Copyright © 2016 Kandimalla, Dash, Kalita, Choudhury, Malampati, Kalita, Kalita, Devi and Kotoky. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: Raghuram Kandimalla, email@example.com Jibon Kotoky, firstname.lastname@example.com


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Posted by obird on Apr 30, 2015 in Recent Work | Comments Off on Excavation Services in Napanee, Kingston and Area
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Win a bib for the 2020 Virgin Money London Marathon and a $1,500 travel allowance courtesy of Zwift and New Balance.
Hyatt and Lindblad Expeditions Officially Launch Partnership to Deliver Wondrous New Travel Experiences
See all of the amazing places you can explore with World of Hyatt and Lindblad Expeditions!
Hyatt Regency Cartagena – The Best Hotel Run-Around Ever!
You know a hotel is special when the staff plan a 5K run in your honor!
Fitness Center Review and Safety: Hyatt House, Jersey City, New Jersey
A well equipped gym with new equipment but I tripped and hurt myself over an uneven surface at the entrance
NYC Marathon: United Offers Free Medal Engraving and a Special Menu For Marathon Runners
United Airlines a leading sponsor of the NYC Marathon has created a new menu with the runners in mind!
New York City Marathon – The Last 10 Miles
Running the last ten miles of the New York City Marathon course a week or two before Marathon Sunday is a tradition among local New York City runners.
The Sporting Club – Fitness Center at the Bellevue Hotel, Philadelphia
This hotel fitness center offers just about everything you could ever want from a fitness center
Fabulous Wearable Art: INKnBURN Fitness Apparel (Great for Travel Too)
INKnBURN creates fitness apparel that is not only comfortable and moisture-wicking but is also imaginative, vibrant and conducive to working out.
SPIbelt – Review, Discount and Giveaway!
SPIbelts are perfect personal storage belts for both runners and travelers.
Beer Running – An International Travel Phenomenon
The first beer runners were British officers stationed in Malaysia before World War Two. They would get together once a week to 'run off the previous weekend's excesses.' And so began the Hash House Harriers.
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Madame Nhu
Ken Burns has produced one of the best narratives of the Vietnam War that the last two American generations have no recollection. I lived through it and I worked on military marketing accounts in the aftermath of the war. In typical fashion, as applies to the previous work of Ken Burns it is spectacular. It is a thoroughly done documentary on how the war was fought in Vietnam, Hanoi, Beijing and how it was fought at home. The Vietnam War tore the country in half and exposed an undercurrent of deeply held but diametrically opposite philosophies in our culture.
Burns interviews living US diplomats and journalists, and he interviews North Vietnamese and Viet Cong combat commanders and UDS combat veterans. During the Kennedy Administration, US and South Vietnamese commanders were reacting to a inexorable rejuvenation of the Communist insurgency especially in South Vietnam where they were out gunned and out number by ARVN and US Forces. Commanders created a method of keeping the peasantry of South Vietnam and the Viet Cong apart.
Introducing one of the worst ideas, or possibly the very worst experiments in social engineering during a conflict. About 85% of the population of South Vietnam lived in the rural woodlands, estuaries and wetlands. So geniuses built over a thousand small villages and reinforced them like military garrisons. That meant Vietnamese who had lived on the same land for centuries were forcibly removed from their homes and forced into military redoubts with a western version of schools, western food all inside heavy guard.
Throughout their history, like the history of all Vietnam, Invaders came and looted and oppressed them from the Mongols, the Ming Dynasty, the Dutch, the Manchus, and the French. The Americans were just another in a long succession of muscular military incursions bristling with aircraft and naval and armor firepower. What exacerbated the problem was that South Vietnamese president Diem was a self-absorbed delusional autocrat. He was despised in the North and in the South. One US ambassador complained that despite all the monetary and strategic support America was providing, Diem was dictating to the Americans. We were kowtowing to this tinplated dictator and his Cruella DeVille wife: Tràn Lê Xuân or Madame Nhu.
When Buddhist Monks were burning themselves alive on the street in front of them, she scorned them contemptuous and glib about Buddhist priests making toast.
The Left and the Right supported the war, like today's neocons on both sides of the aisle. Today the neocons see the Grand Enemy of civilization is Radical Islam. In the 1960s, it was the notion of a monolithic and radical world Communism movement. In the power vacuums in the aftermath of World War II, Communisn did quite well. Some of the largest empires of the world, France, Spain, Russia, and Britain all lost many holdings, and many of these holdings went to the other side and grew into Communist regimes or Socialist regimes. One of the last redoubts of Western mercantile values, Southeast Asia was being wooed and armed by hardline Chinese Communists and by Russian Premier Nikita Krushchev. Also Ho Chi Minh frankly was a charismatic leader highly regarded by poor rural countries in Malaysia and Vietnam and the Philippines. So the US and NATO allies were not doing well against the Warsaw Pact were seeing their influence stall.
The True Believers from Kennedy to Goldwater believed that Vietnam was a price worth paying to keep Maoist Communism from advancing westward towards Cambodia, Malaysia, Laos, Burma and India. It was young full-hearted idealistic American men and women who signed up to keep poor countries from suffering the horror of Communism. Western propaganda rarely mentioned that lots of the countries who went Communist went that way because they chose it. This is not to say that Communism wasn't often foisted upon people as a grinding despotic tyranny. The Khmer Rouge and the Pathet Lao were both perfect examples of Communist insurgencies writ as idealism and more like a heavily armed and organized street gang. About a third of the old Dragnet TV series episodes were Hollywood calumny of Communism.


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NPP > JOURNALS > JAC > Vol. 53 > Iss. 4 (1970)
Volume 53, Issue 4 (1970)
Table of contents and editorial information
R. E. Stevenson
Bridge Across the Image Gap: The St. Paul Story
R. E. Stevenson
Communications—More Than Just Reporting
J. Clayton Herman
Consumerism Meets Agri-Business
Frank Lessiter
An Approach to Communication
Donald W. Poucher
Plan Ahead for Efficiency: This Thing Called Plan of Work
Leo R. Fehlhafer
What's Disney Got That We Ain't?
Bill Paul
Back Matter
The Editor's Corner; Meet ACE Authors
R. E. Stevenson


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And in my view, it's been constructive. In the last few pages, @Gleeok learned about a (major?) problem with integrating ZScript and AS (whatever it is we're doing) he was otherwise unaware of.
I think the real problem with, er, being "constructive", is than nobody knows exactly what's going on. I've discussed various options with lots of people, but nowhere have I seen a decision. I think it'd be helpful if @Gleeok or someone briefly described the current AS work done, and what the current plan for the finished product is, even if tentative. It's hard to discuss AS if we're not all on the same page.


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There are a lot of wacky stories in the Bible and some words way wackier. One of them is "Ebenezer". In the Hebrew, it is אֶ֫בֶן הָעֵ֫זֶר, which is transliterated as . . . you guessed it: Ebenezer. It was originally the name of a place, but then took on another meaning as a stone—a single stone—that was raised by a prophet named Samuel as a monument when he said, "Hitherto hath the Lord helped us.". He did not say those words, of course. That is a translation of what he said, done by some guys who looked at the Hebrew words back between A.D. 1604 and 1611 and then put them into the kind of English that was spoken back then when James I (James One) was the king of England.
I think it is beautiful that way.
But if we were going to say it now, we'd say something like,"God, you have gotten us this far, so we are counting on you to take us all of the way."
Rather than a single stone, multiple stones are sometimes used today if someone wants to raise an Ebenezer (as a reminder of God's provision), probably because it is easier to stack a few or it looks sorta like art or because it is difficult to find a single large stone that looks imposing enough to erect as a monument. And if you did find one it would be quite heavy or maybe tip over or something. So multiple stones are used, and I am using three when I raise mine.
I'll raise it somewhere in the garden, where I'm writing now.
But not now, because I am back home after having prayed in said garden, and that after having looked for a few stones there and then hiding them in a corner for later. That is not my Ebenezer up there in the photograph, it is just three little stones that I put outside my door for a picture (so you'd get the idea). But I have decided to leave them there as a reminder, so I guess it is sorta like an Ebenezer-ette.
There is this old church song with the line "Here I raise my Ebenezer, hither by thy help I've come." I remember as an adolescent singing those words in church and thinking crude thoughts and then looking around with a smirk as if I had uttered some profane, puerile confession and somehow just gotten away with it. Is it just me, or do all young guys think such things when they say some wacky word like Ebenezer? And then you get to raise it. Right there in church. As I said, it is crude, but there it is . . .
When I do finally raise my Ebenezer, it will definitely be in the garden, not right outside my door.
What is the point of all of this prelude? It is this: God has indeed carried us this far. He has made the path straight more than once and He is not going to let some guy like me screw it up. I confess that I am an idiot, and have said and done some things that can easily be described by one word: "stupid." But God himself does not screw around with his plans. He has one for us, and it cannot be thwarted. I may very well be committing the crime of attempted thwarting at times. It is unintended and is not my desire. It is not. There is no excuse other than that I cannot see clearly because my head is in the clouds most of the time. I have said that elsewhere, and I declare it again. My head is over my heels, like others of us, and I just act like a fool.
I am sorry, but I am not likely to change much. I'll grow, I will mature, but I will not be easily altered; I'd be someone else if that happened. So, God help me, I will keep to his path whenever I can see it, and I will just keep going as long as it takes.
For now, it is just me and my Ebenezer . . .


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African grape(redirected from an African grape)
African grape
offensive slang A watermelon. A reference to the stereotype of black people having an affinity for watermelon. (As former African-American slaves grew and sold watermelons following the Emancipation Proclamation in 1863, the fruit became representative of their freedom and was then used as a demeaning stereotype by whites who opposed that freedom.)


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Oiro EDP by Mona di Orio
Year: 2006
Nose: Mona di Orio
Powerful, deep, slap-in-the-head opening of jasmin, heliotrope and immortelle on aromatic cedar woods – rich, carnal and daring, a bit morbid and overhelming with a hint of mind-blowing decadent mess, but that is how "power" scents work. BayKAT's review on Basenotes correctly refers to L'Heure Bleue - different components, but that's the kind of feel. I smell a sort of weird, balsamic-camphor base which somehow amplifies and surrounds this – making the texture almost resemble to the dense power of tuberose-based scents, to which this one in fact smells alike. Perfectly-balanced citrus notes (thank God) refresh the texture. As minutes pass it progressively becomes more and more mellow and sweet, settling down a quite natural powdery floral fragrance, still with a disturbing kind of cloying/rancid note with a weird resinous note floating over like a zombie crow, and an ambery/frankincense subtle base. I frankly can not like this completely, or better say, I am still unsure about when/how to wear this - bold, rich, complex, somptuous, viscious carnal flowers, still with nothing leathery/animalic to provide a darker counterpart: challenging for sure.
Tags: Heure Bleue, Mona di Orio, Oiro
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Tired of throwing away half-used batteries? Wish you knew exactly how much battery life you had left? Worried that rechargeable equals unreliable?
Join us for a special "prime time" webinar about the history of the relationship between the iconic SM57 and the President.
The SM57 and the President of the United States: 50 Years of Service to the White House
The SM57 Instrument Microphone was introduced in 1965. Since the administration of Lyndon B. Johnson, the SM57 has been the microphone of choice for the Presidential lectern.
In this one-hour webinar, Shure Corporate Historian and White House Liaison Michael Pettersen will explain the technical reasons for the use of the SM57 for this critical sound reinforcement application. Behind-the-scenes stories also will illustrate the evolution of the Presidential audio system over the decades. The presentation will be about 40 minutes, allowing 20 minutes for questions at the end.


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Greaves Law - Criminal Law Lawyer
Supplying Criminal Defense Within Robertson Texas!
Call (979) 779-9388!
Did you have an unfair situation with the judicial system? Thinking of in search of a Criminal Law Lawyer? What about a Criminal Law Lawyer? It is imperative to realize that your life is not over. Everybody has slip-ups, but they are problems that can be resolved with an effective Criminal Law Lawyer. Greaves Law is an experienced Criminal Law Lawyer who only addresses the field of defense law allowing him to keep informed concerning the modifications or nuances concerning the legal system. He has already been striving to defend Robertson Texas vicinity offenders for upwards of Fifteen years, and he's had his own law office going back more than 10 years. Contact Greaves Law now to learn exactly why so many in the vicinity of Robertson Texas count on his knowledge as a Criminal Law Lawyer!
From the time it was founded, in 2004, Greaves Law has always been the primary Criminal Law Lawyer in the firm. You won't need to be concerned about finding yourself on trial with a Criminal Law Lawyer you aren't familiar with. Anytime you require a Criminal Law Lawyer and select Greaves Law, you'll get Greaves Law - not some less-prepared attorney from the office who doesn't have the court time. He can personally stay accompanying you from start to finish, fighting to get back your freedom in the vicinity of Robertson Texas!
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Some pursue happiness - others create it.
Wherever you are reading this from: seat back, think about your happiness and just it: feed it, read it, breath it and never fear it: it will be the most powerful instrument you will ever own in your life.
Sometimes we get lost along the way and get influenced by external factors, media being one of them. On another great atempt to market all that is around us, media seemed to have created sereval drive through happy meal packages in which we should find "happy" to go; but from where I'm standing that's just far from what happiness really is to me. We do live on an artificial advertising world, idealized by stereotypes of happiness, which do not correspond to our daily reality, but on the 2nd of july you get to change that and prove them wrong making it right through your own photographs, no touch up's, insted: the real deal. How? Wallpeople, an urban collaborative art project that incentivates people to create a unique moment in an urbarn space, presents this year's project: The Wall Mural of Happiness. This Saturday, over 20 cities simultaneously get to be the real protagonist of a unique exhibition: a wall turned into a improvised open-air museum where everyone can share and show their pictures of their true and personal vision of happiness. This time I won't advise you to CLIP-it, but insted dare you to go out and look for where it will be happening in your own town: Happiness: just do it.
HAPPY CONNECT and my newest friend crush: Mariana Duarte Silva, a force of nature who's purpose on this earth is making it a better place by developing some of the most interesting musical and artistic projects around the globe and bringing them closer to us. She is also responsible for Clipping WALLPEOPLE in Portugal.


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Delwaq: Application of salinity from D-FLOW - Home - Delft3D
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Delwaq: Application of salinity from D-FLOW
Gholamreza Shiravani, modified 1 Year ago.
Delwaq: Application of salinity from D-FLOWPadawan Posts: 65 Join Date: 6/25/16 Recent Posts
Dear D3D-Delwaq community,
could you tell me where is the .sal file from D-FLOW in Delwaq applied? The Salinchlor does not include the salinity from D-Flow. By activating this process, the salinty results will be modelled everywhere zero, it means Delwaq does not include D-FLOW salinity! I realized this salinity of Delwaq is for Transport of salinity as a substance in domain. But where is the salinty from D-FLOW in Delwaq?


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This time of year is always hard on the wallet and if you're like me, this year you're REALLY feeling the pinch. Unfortunately for my bank account, I am a woman who enjoys some of the finer things in life. Also unfortunate- I do not have the monetary assets to afford myself most of these luxuries. So over my years of growing up far less well-to-do than most, I have honed the skill of making small improvements to my life that can give it that little something "extra" and make everything else seem a little more luxe.
For starters, dressing with style doesn't always mean buying brand new luxury clothing and sporting designer handbags. I've found that investing in a few good pieces that can last a lifetime can instantly make the rest of your outfit look expensive. I don't care if you're wearing a $7 t-shirt from Walmart, pair it with a great fitting pair of jeans and a nice watch and you can look like a million bucks. There are certain classic pieces that belong in every wardrobe and if you invest your money correctly and make sure to take care of them, you will never have to buy them again. As I previously mentioned, a great pair of quality jeans that fit perfectly. A classic men's style timepiece, something big enough to be noticed but not big enough to eat your dinner off of. Pawn shops and online consignment stores are chock full of gently used expensive jewelry that you can get for a fraction of the cost. For ladies, the perfect LBD (Little Black Dress)- something that can take you from a job interview to a funeral to cocktails without changing clothes in between. A crisp white button down shirt. Dress shoes and one nice handbag or belt in a color that goes with everything rounds out the complete closet. Make sure to shop places like Beatty Shoes, Ross or Gabe's first for the best deals! They get new shipments 3-6 times a week, and if you're willing to dig, you can find a lot of great stuff there.
The luxe life doesn't start in your closet though. Like charity, it starts at home. Your home is a direct reflection of who you are as a person. What better place to add those little touches and give the impression to all who enter that you have it all together. Start by spending a little extra money on good bedding. A comfy down comforter and 500 thread count sheets can make all the difference between "going to bed" and "OMG I'm never leaving this heaven in the morning." Luckily you can get amazing deals on these items at Ross and TJ Maxx. Drape your beds and living room furniture with plush throws and pillows. One thing I will say to save up your money and invest in for the home-custom framing. You really can tell a difference and a lot of smaller businesses only require you to pay half up front and you can pay the rest off in time. For the kitchen, have a French Press for coffee and always have fresh spa water or sparkling water on hand. (No I don't mean cans of La Croix.) Adding a handful of mint and berries or citrus to a pitcher of regular old tap water will make your guests feel like they're enjoying a night at the Four Seasons and costs you less than $3. In your most intimate of spaces, AKA the crapper, ensure you always have a sturdy plunger and air freshener or matches. You never want to make your guests feel embarrassed. Also, and this is my most sound advice in this article- BUY GOOD TOILET PAPER!! My dad always insisted on Charmin Ultra Strong, no matter how broke we were that week. According to him, "You're never too poor for good sh*t tickets." So ditch the John Wayne tissue (that's rough and tough and don't take crap off nobody) and treat yourself to a little something with an aloe vera finish. No need to thank me later. Also, for your own sake, get a bath tub caddy/tray. I just bought one of these from Target for $21 and it has made every night I've soaked in the tub since feel like a spa treatment. Mine comes complete with a cup holder for wine, a spine to hold up your favorite book or iPad and a flat space on the side for candles or chocolates. My final advice for upgrading your home, if you're of the drinking persuasion, have a complete bar. Whether you have a designated counter space or a rolling tray or a full blown bar in your home, always keep a bottle of the 5 essentials on-hand: vodka, tequila, gin, bourbon/whiskey, and rum. Some staple mixers should always include cola, soda water/tonic, cranberry/orange juice and citrus. Also make sure you have the appropriate glassware that pairs with each beverage. It's also a good idea to keep at least a few light beers and a bottle of red and white wine on hand (pinot noir and pinot grigio seem to be the most crowd pleasing varietals.)
These tips can give you the foundation for living a life that looks AND feels far more expensive and luxurious than it really is. And with all the extra money you're saving you could either invest in your future, or blow on a long weekend away like me! Hey, you cant take it with ya, so spend wisely!