text:
Screening selected medicinal plants for antibacterial activity against Methicillin-Resistant Staphylococcus aureus (MRSA).
IntroductionNature has been a source of medicinal agents for thousands of years. Various medicinal plants have been used for years in daily life to that disease all over the world. Many of modern drugs have been isolated from natural sources are based on their use in traditional medicine. Plants produce a diverse range of bioactive molecules, making them a rich source of different types of medicines. Over 50% of all modern clinical drugs are of natural product origin (Stuffiness and Douros, 1982) and natural products play an important role in drug development programs in pharmaceutical industry.
Staphylococcus aureus is commonly associated with hospital and community-acquired infections. It causes superficial skin lesions such as boils, styes and furunculosis with more serious infections such as pneumonia, mastitis, phlebitis, meningitis, and urinary tract infections; and deep-seated infections, such as osteomyelitis and endocarditis. In 1980s, methicillin resistant cases ascend and vancomycin has been used as a last resort. The prevalence of MRSA isolates in several hospitals in Malaysia in 1996 was even higher at 40% (Rohani et al., 2000). In general the frequency of S. aureus nasal carriage from such settings in Malaysia varies from 45% to 76%. Antibiotics resistance has emerge as a serious problem in Malaysia hospitals and Methicillin-resistant Staphylococcus aureus (MRSA) are reported to be the major pathogen since 1995 (Malaysia Medical Association, 2002).Therefore there is an urgent need to develop alternative antimicrobial drugs for the treatment of infectious diseases.
In order to search antibacterial activity against MRSA strains, four different plants such as Andrographis paniculata (Hempedu bumi), Coleus amboinicus (Oregano), Vitex negundo (Chasteberry) and Hylocereus polyrhizus (Dragon Fruit) were selected. Coleus amboinicus belongs to the family Lamiaceae. It is also known as oregano, or as Karpuravalli inTamil. Synonym for this plant is Coleus aromaticus. Coleus amboinicuss Benth, commonly known as an Indian Borage, is a medicinal plant and several medicinal properties are attributed to this plant in the Indian system of medicine. It is considered to be an antispasmodic, stimulant and stomachic and is used for the treatment of headache, fever, epilepsy and dyspepsia (Morton, 1992). Andro graphis paniculata or known as Hempedu Bumi is from the family Acanthaceae. Andrographis paniculata is also commonly known as Kalmegh is an important medicinal plant, occurring wild in India, and is used both in Ayurevda and Unani system of medicine (Chadha, 1985). The dried herb is a remedy for a number of ailments related to digestion, hepatoprotection, vermicidal, antiacne, analgesic, anti-inflammatory, antibacterial, antityphoid, antibiotic activities, hypoglycemic, besides immune enhancement (Saxena et al., 2000). Vitex negundo is from family Verbanecea and known as Chastetree. The highly dissected leaves of Cut-leaf Chastetree are shaped like Cut-leaf Japanese Maple and were once believed to have sedative effects. The preparation of leaves used in catarrhal fever and applied to sinuses and scrofulous sores. Aqueous extract and oil of seeds possessed anti-oxidant and anti-inflammatory property. Hylocereus polyrhizus a tropical fruit popular in Southeast Asia belongs to the climbing cacti (Cactaceae) family. Dragon fruits reputedly improve eyesight and prevent hypertension. The seeds of the fruit supposed help in controlling blood glucose levels in people with non-insulin-dependent hyperglycaemic conditions and it is also used to treat stomach and endocrine problems.
Materials and Methods
Identification of bacterial strains:
Twenty three clinical isolates of Staphylococcus aureus were obtained from Hospital Besar, Alor Star, Kedah, Malaysia and positive control Methicillin Resistant Staphylococcus aureus (MRSA) strain was obtained from Universiti Kebangsaan Malaysia (UKM). Viability tests for each isolate were carried out by resuscitating the organism in nutrient agar. These clinical isolates were reconfirmed by Gram staining plating on Mannitol Salt Agar which is the selective media for Staphylococcus aureus. Then followed by the specific biochemical test for Staphylococcus aureus such as catalase and coagulase tests were carried out.
Collection of plant materials:
Coleus amboinicus (Karpuravalli), Andrographis paniculata (Hempedu bumi), Vitex nigundo (Nochi), and Hylocereus polyrhizus (Dragon Fruit) were collected from Kedah. These plant materials were identified and classified by a botanist in the Biotechnology Department of AIMST University.
Preparation of plant extract:
Coleus amboinicus, Andrographis paniculata, and Vitex nigundo leaves were collected. The leaves were washed with tap water and allowed to air dry. This was done to reduce the microbial load of plant material due to handling and transportation. The plant materials were dried in shade for three to four weeks. Direct sun light should not be given to the plant materials because the sun light radiation can destroy the active compounds of plant leaves. After drying, the plant leaves were ground finely. This is to increase the surface area of the plant materials when exposed to solvent. The ground plant materials were weighed and stored in separated container. The Hylocereus polyrhizus was cut into four pieces and the skin was removed. The fruits were ground by using blender. Methanol was used for crude extraction purpose using maceration method. The mixture is kept for ninety six hours with periodic stirring. After four days, the mixtures were filtered using Whatman No.1 filter papers. The precipitates were discarded and the supernatant was collected. Each extract was concentrated using rotary evaporator (EYELA, AS). These extractions are primarily used for preliminary screening antibacterial activity.
Antibiotic Susceptibility Test:
Antibiotic susceptibility test was performed to identify the Methicillin Resistant Staphylococcus aureus (MRSA) from these 23 clinical isolates. Mueller Hinton Agar was used for this test. Each isolates was spread on the medium and Methicillin antibiotic disc (5[micro]g) was placed on the middle of the culture. It was then incubated at 37[degrees]C for 24 hours. Similar procedures were followed for oxacillin and vancomycin antibiotics.
Disc Diffusion Assay:
The disk diffusion (Kirby-Baurer) technique, which is of the recommended standards of the National Committee for Clinical Laboratory Standards (NCCLS), was used for antimicrobial test. An overnight suspension culture of MRSA was standardized according to no. 0.5 McFarland Standard (1 x [10.sup.8] cell [ml.sup.-1]) and spread on Mueller-Hinton agar (MHA) media. All the sterilized discs were placed on agar media. The disc was impregnated with the CACM leaves extract at 100 mg. [ml.sup.-1]. Methanol was used as negative control and standard antibiotic vancomycin (30 [micro]g/ml) as positive reference to determine the sensitivity of the strain. The inoculated plates were incubated at 37[degrees]C for 24 h (Karaman et al. 2003). The antimicrobial activity was evaluated by measuring diameter of the inhibition zone (mm) around the disc.
Determinination Minimum Inhibitory Concentration (MIC):
The inoculum of MRSA was prepared from 18 h Nutrient broth culture and adjusted to no. 0.5 McFarland standard turbidity. The CACM leaves extract was dissolved in methanol. Then serial twofold dilutions of test samples were prepared in a concentration range from 6.25 to 400 [micro]g/ml in 10 ml sterile test tubes containing Mueller Hinton (MH) broth. The positive control was MH broth with standard reference antibiotic and inoculums and negative control was the MH broth and inoculums. The test tubes were vortex gently to mix the content and incubated at 37[degrees]C for 24 h (Roland et al. 2007). The MIC value was determined as the lowest concentration of CACM leaves extract in the broth medium that inhibit the growth of MRSA.
Determination Minimum Bactericidal Concentration (MBC):
For the determination of minimum bactericidal concentration (MBC), one hundred microliter of the culture from the diluted sample of MIC assay was transferred into nutrient agar. The liquid was spread plate and incubated at 37[degrees]C for 24 h. The MBC was recorded as the lowest concentration of the extract that gave complete inhibition of colony formation for the test bacteria at latter cultivation (Ronald et al., 2007).
Time-Killing Profile of MRSA:
The time-killing of MRSA with half, one and two fold MIC concentration over time was plotted to asses the bactericidal effect. The CACM leave extract was added to an aliquot of 25 ml nutrient broth in an amount which would achieved the concentration of 0.5, 1 and 2 fold MIC. After 18 hours, MRSA culture randomly chosen was harvested and adjusted by using no. 0.5 McFarland standard. One milliter of adjusted culture transferred into CACM leave extract and nutrient broth and nutrient broth alone (control). The samples were incubated on the water bath at 37[degrees]C. After the addition of the adjusted culture, 1 ml of the test sample was transferred onto nutrient agar and spread plates. The plates were incubated at 37[degrees]C for 24 h and determined the CFU [ml.sup.-1]. The growth of MRSA was measured every 4 h for 48 h (Yoga Latha et al., 2007).
Brine Shrimp Toxicity Assay:
Brine shrimp eggs, Artemia salina, hatch in artificial seawater. Thus artificial sea water was prepared by dissolving 38g of salt in 1 L of distilled water. A total of 0.5g Artemia salina were transferred into artificial sea water and incubated for 24 hours under room temperature with oxygen supplement and light intensity. MRSA extract was prepared in various concentrations for toxicity test. One milliliter from MRSA working solution was aliquot and dissolved in 10 ml of 2% dimethylsulphoxide (DMSO) to obtained 10 mg/ml concentration of extract. Then 1 ml of diluted extract was aliquot and transferred into 10 ml of artificial sea water to obtain 1 mg/ml. This method was followed by two fold serial dilution until to 0.008 mg/ml. 15 Artemia salina was transferred into each dilute extracts and incubated for 12 hours in light intensity condition. The mortality rate of Artemia salina and lethal concentration at 50% ([LC.sub.50]) were calculated (Carballo et al., 2002).
Bioautography assay:
Thin Layer Chromatography (TLC):
TLC was performed on a silica gel aluminum plate (10 x 20 cm, Silica gel 60 [F.sub.254], Merck) to fractionate active compound of 100 mg/ml CACM leave extract. The TLC plates were cut into 2 cm x 8 cm and dried into oven at 90[degrees]C for 10 minutes. This is to activate the TLC plates by absorbing the moisture content from the plates. Then the extract was spotted on the bottom of the plates. Then the TLC plates were transferred into beaker which contains separating solvent. The plates were placed in ascending direction in a beaker with chloroform: methanol: distilled water (25:3.5:0.5) solvent as mobile phase. The plates were observed under UV light. The separated spots were marked and the [R.sub.f] value was calculated.
Bioautography technique:
Bioautographic method is another technique that is used in screening antibacterial activity. Bioautographic method is basically to localize the antibacterial compound from crude extract into chromatogram. This is a supportive and quick search method for antibacterial activity compound. The TLC plates that were used to separate active compounds in CACM leave extract were directly mounted on to seed culture medium. The culture plate was kept for five hour in room temperature to make sure separated compound diffuse into the medium. Then the TLC plates were taken out and incubated at 37[degrees]C for 24 hours (Paul Cos et al., 2006.)
Statistical analysis:
Each experiment was carried out three times. The data in this study was computed and expressed as mean [+ or -] standard deviation and also in SEM. Statistical analysis (ANOVA) was performed using GraphPad Prism Software version 4.0. P [less than or equal to] 0.05 was considered statistically significant for all comparisons.
Results and discussion
The methanol yield extracts of the Androgrphis paniculata, Coleus amboinicus, Vitex nigundo and Hylocereus polyrhizus were presented in Table 1. The highest quantity of material was extracted from Coleus amboinicus (5.28%), while the lowest quantity was obtained from Hylocereus polyrhizus (1.22 %). In Masoko's antifungal analysis, it was revealed that organic solvent methanol is the best extractant in quantitatively and qualitatively. The methanol extract is competent to yield greater amount of extract and also contain higher polar compounds and tannins (Masoko et al., 2007) which are generally act as antimicrobial agents.
Identification of Bacterial Strains:
Hospital isolates bacterial strains were identified as MRSA using standardized microbiology technique. The colonial appearance of 23 nasal cavity clinical isolates was identical and typical. All colonies were golden yellow, opaque, circular, smooth and creamy colour. Gram-staining revealed that Staphylococcus aureus isolates are Gram positive cocci in grape like clusters (Fig. 1). Qualitative biochemical coagulase test confirmed that all the isolates are positive for S. aurues. Among these 23 isolates, only 4 isolates showed a positive catalase reaction and the remaining 19 were catalase negative (Table 2). Recently few catalase-negative Methicilin Resistant Staphylococcus aureus strain have been reported to cause outbreaks in Brazilian hospitals (Del' Alamo et al., 2007). Therefore 19 clinical isolates of catalase-negative Staphylococcus aureus were used for antibiotics susceptibility test to identify methicillin resistant strains.
Antibiotic Susceptibility Assay:
Antibiotic susceptibility test was performed to identify the MRSA strains. In general nosocomial MRSA is multi-drug resistant. During 1944, the first reports of penicillin-resistant S. aureus had appeared and today all strains of S. aureus are resistant to natural penicillins, aminopenicillins, and antipseudomonal penicillins (Chambers et al., 2001; Carballo et al., 2002). Therefore the antibiotic test was done for the ten catalase negative S. aureus with methicillin (5 [micro]g) and oxacillin (1 [micro]g). Only ten nasal cavity clinical isolates shows positive results to methicillin and oxacillin antibiotics. Vancomycin antibiotic is used to treat MRSA infection (Stevens et al., 2002). Therefore vancomycin 30 [micro]g antibiotic discs were used to test against all the ten isolates which are showing MRSA positive. As a result, all the ten isolates were susceptible to vancomycin antibiotics (Table 3).
Disc Diffusion Assay:
The antibacterial activity of four different plants including Andrographis paniculata, Coleus amboinicus, Vitex nigundo and Hylocereus polyrhizus methanolic crude extracts were evaluated through standard disc diffusion method. In the disc diffusion technique, a reservoir containing the test compound at a known concentration is brought into contact with the inoculated medium. The diameter of the clear zone around the reservoir (inhibition diameter) is measured at the end of the incubation period. In order to enhance the detection limit, the inoculated system is kept at lower temperature for several hours before incubation to favour compound diffusion over microbial growth. Thereby it increases the inhibition diameter efficiently (Paul Cos et al., 2006). After 12 hours incubation period, CACM leave extract conferred prominent results compared to Andrographis paniculata Vitex nigundo and Hylocereus polyrhizus crude extract. Clear and larger inhibition zones were formed surrounding the CACM leave extract (Table 4). This is the first time whereby CACM leave extract has been used for the evaluation of antibacterial activity against MRSA strains. Other two plants methanolic extracts such as Vitex nigundo and Hylocereus polyrhizus does not show any better inhibition zones. As CACM leave extract gave prominent results compared to other plants extracts, therefore the same plant extract was chosen for further tests against MRSA strains.
Broth Dilution and effect of CACM leave extract against MRSA strains:
Table 4 shows MIC of CACM leave extract against 11 MRSA clinical isolates. MIC is defined as the lowest concentration that is able to inhibit any visible microbial growth and below that concentration which, there is no further inhibition. The minimal bactericidal concentration (MBC) is determined by plating-out samples of completely inhibited dilution cultures and assessing growth (static) or no-growth (cidal) after incubation (Paul Cos et al., 2006). MIC of this extract is 200[micro]g/ml. However, when this concentration was lower, the broth containing the extracts was turbid. Growth of a MRSA strain in broth culture with CACM leave extract was assessed by determining the optical density at 540 nm in each four hours interval for 48 hours. The time-killing profile of MRSA in the presence of extract was shown in Figure 2. The growth profile of MRSA was inhibited at 4 h for MIC and two fold MIC (400 [micro]g/ml). The log phase of MRSA was reduced from 28h to 20 h in half fold MIC value (100 [micro]g/ml). This exhibits that the extract decelerated the reproduction of MRSA. The time killing profile of MRSA with CACM leave extract prove that the effectiveness of the extract within 48 h.
Brine Shrimp Toxicity Assay:
The brine shrimp lethality assay was proposed by Michael et al. 1956; Carballo et al., 2002. The results revealed that [LC.sub.50] of CACM leave extract was 182 ug/ml. In primary toxicity evaluation of plant extracts by brine shrimp lethality bioassay, [LC.sub.50] values lower than 1000 [micro]g/ml are considered bioactive (Meyer et al., 1982). Therefore the extract consists relatively high amount of bioactive constituents. Besides this, the extract also might consist of bioactive compound which is cytotoxic to Artemia salina. Therefore for more accurate result for toxicity assay, animal models and isolate pure compound should be further evaluated.
Bioutography Assay:
Bioautography localizes antimicrobial activity on a chromatogram using three approaches such as direct bioautography, contact bioautography and agar over lay bioautography (Hamburger and Cordell, 1987, Rahalison et al., 1991, Paul Cos et al., 2006). Separated fragments were visualized under Ultraviolet (UV) light and the [R.sub.f] values were calculated. The TLC plates had eleven separated spots. Contact bioautography method was used to localize antibacterial activity compounds of CACM leave extract from TLC plates to an inoculated agar plate through direct contact. Two separated spots which corresponds to [R.sub.f] values 0.347 and 0.292 revealed inhibition zone on the culture medium (Fig. 3). Therefore it can be presumed that these two spots may have antibacterial activity compounds in CACM leave extract.
MRSA strain growth curve in different concentration of Coleus ambonicus extract
MRSA is important nosocomial pathogens in Malaysia and several recent reports have shown that its prevalence as an endemic nosocomial pathogen is increasing. Methicillin-resistant Staphylococcus aureus (MRSA) as a hospital pathogen has presented many clinical problems in the University Hospital, Kuala Lumpur, Malaysia since 1978. It was noted that the incidence of MRSA among S. aureus isolated from hospital inpatients had increased from 11.5% in 1979 to 18.8% in 1985 (Rohani et al., 1999).
The identification of hospital isolates bacteria strains is very important to confirm the presence of MRSA infection in patients. During the identification coagulase test was carried out. This is an important test to differentiate S. aureus from other species especially from Staphylococcus epidermidis. S. aureus produces coagulase, an enzyme like protein that clots oxalated or citrated plasma which is generated in both esterase and clotting activities. Generally catalase test is used to differentiate Streptococcus from Staphylococcus. However now it is difficult to verify this hypothesis because there are few cases on catalase-negative MRSA infections. First case of catalase-negative MRSA was reported by Xavier and his colleagues in 2001. This catalase-negative MRSA specimen was isolated from a 70-year-old woman, suffering from an Alzheimer's disease (Xavier et al., 2001). Therefore catalase-negative S. aureus strains were chosen for identification of MRSA strains and warrant for the further antibacterial screening. The hospitals isolates catalase negatives S. aurues exhibit methicillin and oxacillin resistant. Resistance to these drugs occurs because of the acquisition of genes that encode drug-inactivating enzymes known as [beta]-lactamases. The expression of the mecA gene encoding low-affinity penicillin-binding protein PBP2a confers resistance to other [beta]-lactams in addition to methicillin. Besides that, transformation of a SCCmec type I element into S. aureus strains yielded highly oxacillin-resistant (Utsui et al., 1985; Louis, 2006). The MIC and time killing profile reveal the effectiveness of the CACM leave extract MRSA clinical strains. The MRSA growth inhibition is may due to the active compound from the extract. This statement is pretty true due to the comparison of the control MRSA growth for 48 h with the extract. The reduction in the growth rate may due to the restriction of cell division in the culture. Therefore again it is proven the effectiveness of the extract against MRSA strain. The brine shrimp assay is based on the ability to kill laboratory cultured Artemia salina naupii. The Artemia salina lethality assay is considered a useful tool for preliminary assessment of toxicity. The brine shrimp assay is very useful tool for the isolation of bioactive compounds from plant extracts. The method is attractive because it is very simple, inexpensive and low toxin amounts are sufficient to perform the test in the microwell scale. In addition, this method is rapid, simple, reproducible and economical. A wide variety of biologically active chemical compounds, in particular cytotoxic agents are toxic to brine shrimps. Bioactive compounds are nearly always toxic in high concentrations and as toxicology can be described as pharmacology at higher doses. This premise has been applied to the screening of medicinal plant extracts in the brine shrimp toxicity test. The TLC two blue colour spots exhibits inhibition area in the biouautography assay. Under UV light these two relative spots expressed blue colour. Generally flavonoid derivatives express blue colour under UV light. Besides that, Alcaraz and colleagues (2000) reported that specific growth rate of MRSA strains linearly decrease with the increase in the concentration of flavonoids in the culture medium.
Therefore, it can be assumed that the antibacterial activity spots in TLC plates may be flavonoids or its derivatives. Further evaluation for the isolation and identification of bioactive compounds from CACM leave extract should be undertaken.
Coleus amboinicus methanolic crude leave extract pronounce promising results against nosocomial MRSA pathogens. In further studies isolation of these two pure compounds which completely inhibit this pathogen should be the next line of research. This can give more promising results on antibacterial activity of Coleus amboinicus methanolic extract against nasal carriage MRSA pathogens.
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Masoko, P., J. Picard, J.N. Eloff, 2007. The antifungal activity of twenty-four southern African Combretum species (Combretaceae). South African Journal of Botany, 73: 173-183.
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Michael, A.S., C.G. Thompson and M. Abramovitz, 1956. Artemia salina as a test organism for a bioassay. Science, 123: 464.
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Roland, N.N., E. Alertia, T. Malange, M. Susan, N.L. Henry, Agnes Malongue, M.N. Lucy, N. Kennedy, W. Clare and M.N.E. Simon, 2007. In vitro anti-Helicobacter pylori activity of extracts of selected medicinal plants from North West Cameroon. Journal of Ethnopharmacology, 114: 452-457.
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(1,2) Sahgal, G., (1,3) Sreeramanan, S., (1,4) Sasidharan, S., (1) Xavier, R., (1,4) Ong, M.T.
(1) Department of Biotechnology, Faculty of Applied Sciences, AIMST University, Kedah, Malaysia, (2) Centre for Drug Research, University Sains Malaysia, 11800 Pulau Pinang, Malaysia, (3) School of Biological Sciences, University Sains Malaysia, 11800 Pulau Pinang, Malaysia, (4) INFORMM, University Sains Malaysia, 11800 Pulau Pinang, Malaysia.
Sahgal, G., Sreeramanan, S., Sasidharan, S., Xavier, R., Ong, M.T.; Screening selected medicinal plants for antibacterial activity against Methicillin-Resistant Staphylococcus aureus (MRSA); Adv. in Nat. Appl. Sci., 3(3): 330-338, 2009.
Corresponding Author: Dr. Xavier Rathinam, Department of Biotechnology, Faculty of Applied Sciences, AIMST University, Kedah, Malaysia.
Email: email@example.com. Tel: 0060164591514
Table 1: The extractive values and % yield of the extracts of the selected medicinal plants Extractive Extracts values (g) % Yield (w/w) Coleus amboinicus (leave) 4.78 5.28 Andrographis paniculata (leave) 7.43 2.33 Vitex nigundo (leave) 7.12 5.01 Hylocereus polyrhizus (fruit) 5.34 1.22 Table 2: Qualitative biochemical and antibiotic tests against Staphylococcus aureus for the identification of Methicillin Resistant Staphylococcus aureus (MRSA). Isolated strains Biochemical tests Coagulase Catalase * PC1 Positive Negative * PC2 Positive Negative * PC3 Positive Negative * PC4 Positive Negative 1 Positive Negative 2 Positive Negative 3 Positive Positive 4 Positive Negative * 5 Positive Negative 6 Positive Positive 7 Positive Negative * 8 Positive Negative 9 Positive Positive * 10 Positive Negative * 11 Positive Negative 12 Positive Negative * 13 Positive Negative * 14 Positive Negative * 15 Positive Negative 16 Positive Negative 17 Positive Negative 18 Positive Negative Isolated strains Antibiotics Methicillin Oxacillin Vancomycin * PC1 R R S * PC2 R R S * PC3 R R S * PC4 R R S 1 S S S 2 S S S 3 S S S 4 S S S * 5 R R S 6 S S S 7 S S S * 8 R R S 9 S S S * 10 R R S * 11 R R S 12 S S S * 13 R R S * 14 R R S * 15 R R S 16 S S S 17 S S S 18 S S S Note: R: Resistant towards the antibiotics; S: Susceptible towards the antibiotics. PC1, 2, 3, and 4 strains resemble as the Positive Control of Staphylococcus aureus from University Kebangsaan Malaysia (UKM). The numerical numbers emphasis the isolates strains from hospitalized respiratory infected patients.* Marked strains were used for further antibacterial screenings. Table 3: Four different plants including Coleus ambonicus (Karpuravalli), Vitex nigundo (Nochi), Andrographis paniculata, Hylocereus polyrhizus (Dragon fruit) were used for the antibacterial screening against MRSA clinical isolates. Isolates Remarks Inhibition zone (diameter, mm) [E.sub.1] [E.sub.2] [E.sub.3] (mm) (mm) (mm) 3 MRSA strain 14 6 6 5 MRSA strain 15 15 6 6 MRSA strain 6 11 6 7 MRSA strain 16 19 6 8 MRSA strain 13 13 6 10 MRSA strain 12 12 6 11 MRSA strain 15 11 12 13 MRSA strain 13 18 6 14 MRSA strain 9 10 12 15 MRSA strain 16 22 6 Isolates Inhibition zone (diameter, mm) [E.sub.4] (mm) Control 3 6 6 5 6 6 6 6 6 7 6 6 8 6 6 10 6 6 11 6 6 13 6 6 14 9 6 15 6 6 Notes: [E.sub.1] Andrographis paniculata (Hempedu bumi); [E.sub.2]: Coleus ambonicus (Karpuravalli); [E.sub.3]: Vitex nigundo (Nochi); [E.sub.4]: Hylocereus polyrhizus (Dragon fruit); Control: Methanol. The inhibition zone more than 6 mm is sensitive towards the extract. Table 4: Qualitative Minimum Inhibition Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of Coleus ambonicus crude methanolic (CACM) leave extract against MRSA clinical isolates. Clinical Isolates Remarks MIC ([micro]g/ml) Extract 3 MRSA strain 200 5 MRSA strain 200 6 MRSA strain 200 7 MRSA strain 200 8 MRSA strain 200 10 MRSA strain 200 11 MRSA strain 200 13 MRSA strain 200 14 MRSA strain 200 15 MRSA strain 200 Clinical Isolates MBC ([micro]g/ml) Extract Vancomycin 3 400 30 5 400 30 6 400 30 7 400 30 8 400 30 10 400 30 11 400 30 13 400 30 14 400 30 15 400 30
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Title Annotation: | Original Article |
Author: | Sahgal, G.; Sreeramanan, S.; Sasidharan, S.; Xavier, R.; Ong, M.T. |
Publication: | Advances in Natural and Applied Sciences |
Article Type: | Report |
Geographic Code: | 9MALA |
Date: | Sep 1, 2009 |
Words: | 4934 |
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One of our succulents has recently developed a white, furry fungus on it's stem. Any suggestions on what it could be and how to fix it?
WANTED: Nerines CAPE PENINSULA
WANTED: Nerines in exchange for different Clivias, indigenous trees, lemon grass or Daylilies. Nicola, Kirstenhof, Cape Peninsula. Tel: 021 701 2847
Buffalo Grass Blues
Hi there, When we moved in there was newly planted buffalo grass in our back garden (south facing), this was about 2 yrs ago and it has died. We think it is because there is little sun, lots of trees and poor soil preparation, i think there is building rubble underneath the soil. We would […]
WANTED: Comfrey Plant Durban
Hi everyone, I am looking to buy fresh comfrey leaves or a comfrey plant in Durban and surrounding areas. Am willing to collect. Could anyone help? Thanks so much, Priya
What is this plant?
I have this plant in my garden and I need to know what it is, it is a very fast grower and already higher than the roof. The leaves grow cylindrical around the stems and it now has green berry like "fruit" in a cluster. Can someone please help?
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- Companion Planting Competition November 16, 2011
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- What is this plant? January 28, 2015


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Walmat FREEZING ALL AMMO...IN ALL STORES!!!
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someone posted this on my fb and he buys his ammo there
A must read this will not only hit the nail on the head but drive it straight through
Wal-Mart vs. The Morons
1. Americans spend $36,000,000 at Wal-Mart Every hour of every day.
2. This works out to $20,928 profit every minute!
3. Wal-Mart will sell more from January 1 to St. Patrick's Day (March 17th) than Target sells all year.
4. Wal-Mart is bigger than Home Depot + Kroger + Target +Sears + Costco + K-Mart combined.
5. Wal-Mart employs 1.6 million people, is the world's largest private employer, and most speak English.
6. Wal-Mart is the largest company in the history of the world.
7. Wal-Mart now sells more food than Kroger and Safeway combined, and keep in mind they did this in only fifteen years.
8. During this same period, 31 big supermarket chains sought bankruptcy.
9. Wal-Mart now sells more food than any other store in the world.
10. Wal-Mart has approx 3,900 stores in the USA of which 1,906 are Super Centers; this is 1,000 more than it had five years ago.
11. This year 7.2 billion different purchasing experiences will occur at Wal-Mart stores. (Earth's population is approximately 6.5 Billion.)
12. 90% of all Americans live within fifteen miles of a Wal-Mart.
You may think that I am complaining, but I am really laying the ground work for suggesting that MAYBE we should hire the guys who run Wal-Mart to fix the economy.
This should be read and understood by all Americans… Democrats, Republicans, EVERYONE!!
To President Obama and all 535 voting members of the Legislature
It is now official that the majority of you are corrupt morons:
a.. The U.S. Postal Service was established in 1775. You have had 237 years
to get it right and it is broke.
b.. Social Security was established in 1935. You have had 77 years to get it
right and it is broke.
c.. Fannie Mae was established in 1938. You have had 74 years to get it right
and it is broke.
d.. War on Poverty started in 1964. You have had 48 years to get it right;
$1 trillion of our money is confiscated each year and transferred to "the
poor" and they only want more.
e.. Medicare and Medicaid were established in 1965. You have had 47 years
to get it right and they are broke.
f.. Freddie Mac was established in 1970. You have had 42 years to get it right
and it is broke.
g.. The Department of Energy was created in 1977 to lessen our dependence
on foreign oil. It has ballooned to 16,000 employees with a budget of $24 billion
a year and we import more oil than ever before. You had 35 years to get it right
and it is an abysmal failure.
You have FAILED in every "government service" you have shoved down our
throats while overspending our tax dollars.
Folks, keep this circulating. It is very well stated. Maybe it will end up in the e-mails of some of our "duly elected' (they never read anything) and their staff will clue them in on how Americans feel.
I know what's wrong. We have lost our minds to "Political Correctness" !!!!!!!!!!!!!!!!!!
Someone please tell me what the HELL's wrong with all the people that run this country!!!!!!
We're "broke" & can't help our own Seniors, Veterans, Orphans, Homeless etc.,???????????
In the last months we have provided aid to Haiti , Chile , and Turkey ..And now Pakistan ........previous home of bin Laden. Literally, BILLIONS of DOLLARS!!!
Our retired seniors living on a 'fixed income' receive no aid nor do they get any breaks…
AMERICA: a country where we have homeless without shelter, children going to bed hungry, elderly going without 'needed' meds, and mentally ill without treatment -etc, etc.
Imagine if the *GOVERNMENT* gave 'US' the same support they give to other countries. Sad isn't it?
99% of people won't have the guts to forward this.
I'm one of the 1% -- I Just Did
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Found: Early Black Hole 12 Billion Times More Massive Than Our Sun
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Hollywood Studio Caught Producing ISIS Videos
A meteor going 45,000 MPH lit up the sky in western PA. You could see it from Ohio and New York!
America : Freedom to Fascism
Rapper Afroman Caught On Video Punching Woman At Nightclub
USDA Approves First GMO Apple For Planting
US Senator used old photos of Russian troops to push Ukraine war propaganda


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"No provision in our Constitution ought to be dearer to man than that which protects the rights of conscience against the enterprises of the civil authority."
–Thomas Jefferson
The UC Hastings student CLS chapter welcomed all members of the university community to participate in its activities but required its officers and voting members—who spoke on its behalf, voted on its policies and programs, and led its Bible studies—to share and abide by the group's core beliefs.These beliefs included being Christian and abstaining from sexual activity outside of marriage.UC Hastings decided at the beginning of the 2004 school year that CLS's voting membership and office-holder requirements violated the religion and "sexual orientation" provisions of its Policy on Nondiscrimination.UC Hastings then denied CLS "Registered Student Organization" (RSO) status.
That same year, approximately sixty RSOs—organized around diverse interests in politics, religion, culture, race, ethnicity, and human sexuality—existed on campus.However, the CLS student chapter became the only group ever denied RSO status at UC Hastings.
CLS then sued, claiming that UC Hastings violated its constitutionally protected rights of free speech, expressive association, free exercise of religion, and equal protection of the laws. Unfortunately, CLS was denied relief by the federal courts, including the U.S. Supreme Court, because of the specific nature of the policy at UC Hastings, which allegedly required student groups to accept all students, regardless of their status or beliefs.The Court concluded that public universities may override a religious student group's right to determine its leadership only if it denies that right to all student groups.
The decision in CLS v. Martinez, 561 U.S. __ (2010) could have a damaging effect on the religious liberty of all students attending public colleges and universities.The decision puts many other student groups across the country at risk and leaves room for absurd scenarios, such as requiring CLS to allow atheists to lead its Bible studies.Recently, a similar policy at private Vanderbilt University forced the school's Catholic student group off campus because Vanderbilt Catholic requires that its leaders be Catholic (although it allows anyone to be a member of the group).
Is our most cherished freedom truly under threat?Among many current challenges, such extreme "nondiscrimination" policies deprive students of the right to exercise freely and fully their religious beliefs.Religious liberty is not only about our ability to go to Mass on Sunday or pray the Rosary at home.It includes our ability to gather with other members of our faith outside of church and reinforce our beliefs within a group setting.Without religious liberty properly understood, all Americans, including students on campus, suffer.
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Easter devotions are here! Sign up for our Lent and Easter email devotions.
33:7 the waters of the sea. The creator is pictured poetically as exercising command over the waters of chaos (Ps. 18:16 note). There may also be a poetical allusion to the song of Moses and Israel at the Red Sea (Ex. 15).


text: Hands On: Dragon Ball Z: Infinite World – Gah! Too Many Colons! AJ GlasserProfileFollowUnfollow AJ GlasserFiled to: Impressions Dragon Ball Z: Infinite World Atari dimps 9/12/08 11:00am 9/12/08 11:00am EditDelete InviteInvite manuallyPromoteXDismissXUndismissBlock for KotakuHideShare to KinjaShare to FacebookShare to PinterestShare to TwitterGo to permalink Kids today don't know their anime – it's all Naruto this and One Piece that. And don't even get me started on Death Note. Remember when Dragon Ball Z was the only anime we got on Cartoon Network? And it had that really lousy dub with the "It's over 9000" line that will live in meme infamy? There was also a time when Dragon Ball Z: Budokai was the only anime fighting game we had in North America. You remember it – that chunky PS2 game with the taunt button and the airborne battles. It got mixed reviews, but sold well enough to justify several sequels, including the reincarnated Burst Limit on next-gen consoles. And now, because developer Dimps has moved onto bigger and better things Sonic Unleashed Wii level designing, they've decided to come back with a last hurrah – Dragon Ball Z: Infinite World.A pastiche of all the Budokai games (even the the PSP Shin Budokai titles), Infinite World features the best of the combat features and the frenetic fighting style the series is famous for. So finally we get the aura dash and the doge and weave all in the same game – plus the capsule system has returned, so you can customize your characters' move set. They even got the original Japanese voice actors and the second-gen dub voice actors to reprise their roles. Thought I don't know how happy I am about that – I liked the first generation of dubbers, when Piccolo sounded like a chain smoker and Frieza didn't have laryngitis. In addition to versus mode, the story mode (called Dragon Mission) also features minigames that play more like platformers . I got to watch Goku tear down Snake Way towards King Kai's mini-planet, trying to pick up power-ups without falling off into hell home for infinite losers. There are also race-attack and time-attack levels and a button-mashing exercise in the hyperbolic time chamber. Dragon Ball Z: Infinite World has over 40 characters (not counting evolved forms) and will the plot of the anime from Raditz's arrival through the animated abortion that is GT. Seriously, how many of you have seen the anime? I'm pretty sure that's why Dimps went all-out with story cutscenes – they're assuming the core audience is too jacked up on everyone's favorite orange-clad ninja to even know who Vegeta is. The game's out this November. Here, have some screenshots: 164Reply 164Discuss


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Ok so i decided to do my first batch of cider and im using the mangrove jacks pear cider. When i initially started my batch i took a gravity reading of 1.010, about 5 days later after it had seemed the bubbling in the airlock had come to an end i took another gravity reading of 1.020, THEN we opened the lid and added the pear flavoring giving it a quick stir and closed the lid like the directions said, However this morning (only two days later) i just another gravity reading and its at 1.032!!! whats going on?! also I noticed the water in my airlock doesnt look right. All the water is in the first chamber (my airlock is the kind that there is two little chambers side by side connected at the bottom by a smaller tube). when it is fermenting the air pressure from the inside pushes the water into the second chamber, when it is nearly finished its kinda balanced between the two and now its as if the pressure from outside my fermenter is pushing back INTO my fermenter AND i have also heard it bubble a couple times the last couple days. Is my fermenter sucking air from outside back in?!?! is my cider completely trashed?
Did you check to see if your hydrometer is working right? if you put it in a glass of water and it doesnt read 1.00 you know the issue. The only other thing I can think of is that your having a LOT of water evaporate from your cider making the gravity go up... That or someone's playing a prank on you and throwing things into your cider.


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Bebi water
Started by
, Jun 13 2012 09:04 AM
49 replies to this topic
#1Posted 13 June 2012 - 09:04 AM
Just looking at the latest woolworths catalogue and they have a product called 'Bebi Infant suitable water'.
It also has a ready to use infant teat so all you have to do is add formula or just give them water. #2Posted 13 June 2012 - 09:11 AM
Its a good marketing ploy for those who think its too hard to prepare a bottle of water.
I certainly won't be buying one... #3Posted 13 June 2012 - 09:15 AM
They come in fruit juice too
#4Posted 13 June 2012 - 09:16 AM
Wow, imagine the mark up on it!
Sounds like when they started selling pre-cut fruit. #5Posted 13 June 2012 - 09:19 AM
Should have put it is $1.95 in the catalogue, normal price will be $2.95. For a 250ml bottle.
#6Posted 13 June 2012 - 09:27 AM
it's a convenience. i can see it coming in handy for people in an emergency (ie a bottle has leaked/no boiled water available), much like formula stick packs/'poppers'.
#7Posted 13 June 2012 - 09:31 AM QUOTE (livvie7586 @ 13/06/2012, 09:27 AM) <{POST_SNAPBACK}> it's a convenience. i can see it coming in handy for people in an emergency (ie a bottle has leaked/no boiled water available), much like formula stick packs/'poppers'. Exactly what I was thinking! You wouldn't buy it to use every day, but if you get caught out at home or if you're out and have no other option than to buy a tin of formula and a bottle of water... If DD ends up bottle fed then I would consider having one as an emergency. #8Posted 14 November 2012 - 03:36 PM
I'll be boycotting this product on the basis that they actively market fruit juice to babies in a bottle with a teat. They have a Facebook page where they are pretending to be open to hearing issues from concerned parents, but when someone posts a comment they don't like, or a link to nutrition advice from WHO and NHMRC, they are actually banning people and deleting their posts!
Edit: I have deleted my comment about Bebi just selling the water used to mix formula. Because I have never used formula, I didn't release that it's not just the water that needs to be sterile, the formula has to be heated to 70ºC and then cooled to be sterile... so unless parents are able to heat the bottle after mixing, this could be dangerous as well! I believe you can buy tetrapacks of premixed formula that is safe for babies and would be far safer than using this product! I've also posted a link to this statement by the Dental Health Association Edited by MelbChick, 15 November 2012 - 03:34 PM. #9Posted 14 November 2012 - 03:38 PM QUOTE (MelbChick @ 14/11/2012, 01:36 PM) <{POST_SNAPBACK}> I'll be boycotting this product on the basis that they actively market fruit juice to babies in a bottle with a teat. They have a Facebook page where they are pretending to be open to hearing issues from concerned parents, but when someone posts a comment they don't like, or a link to nutrition advice from WHO and NHMRC, they are actually banning people and deleting their posts! Beats me why they couldn't have just sold the sterile water product for mixing formula and left it at that +1 #10Posted 14 November 2012 - 03:41 PM
i can see the use of a the product...
there were a few times with boys boys that i had to purchase a bottle and bottle of water because i had forgotten to pack a bottle... #11Posted 14 November 2012 - 03:47 PM
I noticed this last week or the week before on one of those morning shoes as I was running out to work. Missed half of it but it was like an infomercial segment.
Only based on the little bit I did see it's the biggest load of crap I have come across in a while. Still to this day I do not buy juice or cordial and can not see why it should be part of any childs diet who is still needing a teat! #12Posted 14 November 2012 - 04:15 PM QUOTE I'll be boycotting this product on the basis that they actively market fruit juice to babies in a bottle with a teat. They have a Facebook page where they are pretending to be open to hearing issues from concerned parents, but when someone posts a comment they don't like, or a link to nutrition advice from WHO and NHMRC, they are actually banning people and deleting their posts! Beats me why they couldn't have just sold the sterile water product for mixing formula and left it at that I think it is terrible and the juice is just wrong. If they did any research at all they would find very quickly that babies/infants/toddlers should NOT have juice in their bottles as per recommendations from Paediatric Dentists. Baby Bottle Tooth Decay anyone? #13Posted 14 November 2012 - 04:19 PM
I am another boycotting them based on their juice products. Disgusting company.
#14Posted 14 November 2012 - 04:19 PM
sounds like another w**k product
#15Posted 14 November 2012 - 04:23 PM
I thought I saw this product on the bus today. I was wondering what the mum next to me was doing unwrapping the bottle and snapping off the tip...
Thanks for clarifying what it actually is. Won't be buying either variety either.... #16Posted 15 November 2012 - 03:31 PM
This statement has been released by Dental Health Services Victoria
Babies Don't Need Bebi Fruit Drinks #17Posted 16 November 2012 - 10:03 AM
They are certainly copping it on their Facebook Page.
#18Posted 16 November 2012 - 10:03 AM
Ooops double post
Edited by Sambambino, 16 November 2012 - 10:03 AM. #19Posted 16 November 2012 - 10:13 AM
They were handy when we travelled with a baby recently to a place without potable water. I took half a dozen of them.
I saw the juice and just said wtf to myself. #20Posted 16 November 2012 - 10:36 AM QUOTE (MelbChick @ 15/11/2012, 04:31 PM) 15081738[/url]'] This statement has been released by Dental Health Services Victoria Babies Don't Need Bebi Fruit Drinks Thanks for the link. I am disgusted at this product. Why is woolworths even stocking this? Just evil. #21Posted 16 November 2012 - 01:26 PM QUOTE (~*Twilight~Zone*~ @ 16/11/2012, 02:21 PM) <{POST_SNAPBACK}> Say what? It that just these bebi bottles or in normal formula usage? No you don't boil formula. You don't have to sterilise even. It is just reccommended hot soapy water to wash and air dry. Some people still like to sterilise their bottles. But as most peoples kitchens arent sterile it doesn't really matter. You use cooled boiled water to make up the bottles. #22Posted 26 November 2012 - 10:52 PM
This product looks pretty bad
I think one of the juices is white grape, seriously it sounds like white wine! I wonder if it is the same people who make alcho-pop type drinks? #23Posted 26 November 2012 - 11:46 PM
The product is bad but seriously? You really think grape juice is bad? To turn into wine it needs to ferment.
No way would they put wine in a product designed for babies. #24Posted 26 November 2012 - 11:56 PM #25Posted 27 November 2012 - 05:53 AM QUOTE (Avocado @ 16/11/2012, 11:36 AM) <{POST_SNAPBACK}> Why is woolworths even stocking this? Just evil. Probably because people are buying it. Maybe evil people. This guy perhaps QUOTE I am disgusted at this product. Here's the thing, if you don't like the product then don't buy it! 1 user(s) are reading this topic0 members, 1 guests, 0 anonymous users |
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The splash from rain hitting a windowpane or printer ink hitting paper all comes down to tiny droplets hitting a surface, and what each of those droplets does.
A new study from a pair of Cornell University researchers has found describing a portion size can affect eating habits in the same way smaller portions can reduce food intake.
Leading higher education marketing firm mStoner announces the launch of the new Advancement Live channel on the Higher Ed Live network.
- A pivoted catch designed to fall into a notch on a ratchet wheel so as to allow movement in only one direction (e.g. on a windlass or in a clock mechanism), or alternatively to move the wheel in one direction.


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Best, Worst Warriors Draft Picks Since 2000
by Zach Buckley
As it currently stands, the Golden State Warriors hold no picks in Thursday's NBA draft. That's perhaps the biggest sign of the franchise's recent turnaround, as the league's annual ... Read More »
Arenas Trolls Stephenson Big-Time
by Tristan Thornburgh
Former Washington Wizards point guard Gilbert Arenas showed no mercy as he mocked Indiana Pacers center Roy Hibbert after his poor Game 1 performance against his former team in the Eastern C... Read More »
Arenas Compares John Wall and Bradley Beal to Himself and Larry Hughes
by Grant Hughes
Gilbert arenas knows a thing or two about exciting backcourt duos, so he's probably qualified to comment on the Washington Wizards ' current tandem of John Wall and Bradley Beal... Read More »
Nick Young Not a Fan of Gil's Selfie
by Tristan Thornburgh
Former NBA star Gilbert Arenas recently posted his first selfie on his Instagram account with the caption "first selfie lmao." Lakers swingman Nick Young, Arenas' former teammate on the Washing... Read More »
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Product Description
Available while supplies last.
This Giagni Single Lever Lavatory Faucet is offered in a polished chrome finish.
Single Lever Lavatory Faucet
- Mounts in 4-in spread or single hole
- AB 1953 Low Lead Compliant
- 1 1/2-in maximum deck thickness
- 1/2-in female supply with 1/2--in to 3/8-in adapter
- Solid brass construction and ceramic disc cartridges ensure years of reliability
- Optional deck plate
Finish: | Polished Chrome | |
Materials: | Solid Brass | |
Weight: | 5 Pounds | |
Listing: | Quick Ship, ADA, WaterSense | |
Faucet Type | Vessel |
Dimensions & Weights
Height: | 8 inches |
Width: | 3 inches |
Liters per Minute (Max): | 5.69L |
Gallons per Minute (Max): | 1.5gal |
Spout Reach: | 5 inches |
Spout Height: | 4¼ inches |
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Group # | 1453908 |
Category | Bathroom Faucets - 456 |
Style | Transitional - 15 |
Sales Rank | 253 |
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-or- Register at the Citadel
Lady Celia, Tempest Queen
NPCs (Major) (Political)
's comment on 2005-01-28 10:25 AM
Add the link to Mytrian the wise and perhaps add a few more details so people will not be so confused.
Try to anticipate the questions your reader might have and either answer them or gently nudge them in the right direction so they can make their own decision.
Go to Comment
The Lonesome Village
Locations (Forest/ Jungle) (City)
's comment on 2003-07-30 07:55 AM
A neighbor to Asylum or a smaller scale version, easy to put into a campaign.
What causes one to be lost? More specifically, why are there so many that someone had to make a special place for them?
Was there a great Evil that came across the land?
If monsters were responsible, then a number of ex-adventuring types would be there. Perhaps they are the mcguffin that brings you there.
What noble ruler (or was it a church) that ran/ runs the hospital/ asylum? And why do they run it? Is it guilt that motivates them or is it altruism?
Questions, Question, and more questions. Nice concept and easy to impliment
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The Forbidden Jungle
Locations (Area) (Forest/ Jungle)
's comment on 2005-11-05 05:50 PM
A handful of votes and 1713+ views and I am the first comment? tsk. tsk.
Go to Comment
Mirra Soothe
NPCs (Scenario Based) (Religious)
's comment on 2005-11-05 05:58 PM
The voting population did not comment on this, nor did the over one thousand other viewers this post has had.
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The Cliff and Valley
Plots (Mystical) (Encounter)
's comment on 2005-11-06 06:48 PM
It should of had more votes in the past. I don't know why they didn't transfer. This is what I voted a long time ago.
Go to Comment
Locations (Forest/ Jungle) (Country/ State)
's comment on 2005-11-05 05:47 PM
This post has had 1518 view and eight plus votes, yet nobody wants to comment?
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The Tome of Klorak
Plots (Discovery) (Single-Storyline)
's comment on 2003-07-23 02:25 AM
That is why people are not allowed "in and out". If the Mad learned that you can get out, they will try... in droves. (Did we mention that they are superhumanly strong and immune to pain?) That is why people are not allowed in or out. That is why you have to sneak in and out.
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The Tome of Klorak
Plots (Discovery) (Single-Storyline)
's comment on 2004-09-18 09:15 AM
Go to Comment
Madness on the road
Plots (Discovery) (Encounter)
's comment on 2004-09-18 09:16 AM
Given an update.
Go to Comment
The Lost Marbles
Items (Other) (Magical)
's comment on 2003-07-17 04:09 AM
Hmmmm... Marbles... Orbs... Corvus?
Go to Comment
The City of Asylum
Locations (City) (Any)
's comment on 2003-07-14 05:38 AM
Good thought. The historical record does not say I (though I always think those on the walls were the only ones that didn't go. I have always throught the material in the walls, some property of the stone. However, I do not know.
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The City of Asylum
Locations (City) (Any)
's comment on 2003-07-15 12:32 AM
I am going to add a few scenarios to link to this setting. I would be mad not to.
Ohhh. and another Orb...
Go to Comment
The City of Asylum
Locations (City) (Any)
's comment on 2003-08-10 02:14 AM
Nope. They are not. Generations have gone by since Asylum went mad (the meaning of the name of the city has changed) and the apparently immortal, near invunerable, and incredibly strong mad people, are still insane waundering about in a mad parody of normal life.
It is said, everyone who enters the city and stays there over long goes mad. Those who go into the city to heal the mad, usually are never seen again. Popular thought is that they go mad themselves before they can do any good or just die there.
If you want there to be expeditions into the city in an attempt to help them in your campaign, feel free. I could see that happening every generation or so, as people forget what happened to the last one OR the organizer thinks they are cleverer than the previous organizer.
Go to Comment
The City of Asylum
Locations (City) (Any)
's comment on 2003-08-10 02:17 AM
Oh and eventually anyone who enters the city will go insane. It all depends on them and the amount of time they spend in the city.
The city guards are very good at spoting "the look" in people's eyes. If they even think you are insane, they will not let you out of the city.
Eventually there will be an overpopulation problem, but so far no.
Go to Comment
The City of Asylum
Locations (City) (Any)
's comment on 2003-08-19 04:49 AM
When playing with any potentially insane person, I will occasionally pull them aside and give them other information about things occuring. You hear shuffling behind you... you feel them looking at you... you hear your friends laughing at you when they think you are not around... you see the monster pulling the strings of the people around you... see CP for more insane things.
Then if they refuse to act upon this information, I pull the rest of the people aside and we run a less than lucid moment. The other players act out the strange reality and the character must react to it appropriately because it is really happening... to them. I then take notes about what the crazy one does and says. We then play out the game, with me telling them what the crazy person does.
Go to Comment
The City of Asylum
Locations (City) (Any)
's comment on 2005-09-15 02:45 PM
plot lines
Occasionally they send the mad from other areas of the land to Asylum. You might encounter an army unit herding some crazies to the city, while taking them supplies.
An interesting idea is to be a young person inside the city, being a little nuts, rather than insane, As you advance in experience you, you advance into madness.
The leader of the country uses Asylum as a prison for political prisoners. "I am the Duke of Ellington! yah, right, another crazy. But, I AM!. Yah. Right. Move along."
An interesting idea is to be a new army officer assigned to watching over the city and realizing how insane the officers are. Are you just nuts? Is that why they sent you here? If they realize you will tell others they are insane, they will stop you... perhaps throwing you into the grand city.
Perhaps you have to risk Asylum to reach the great library, or find a political prisoner, or find the orb of McGuffin (an object of power that is requires to save the city/ world/ temple), or even on an expedition of healers to try and heal the mad.
Go to Comment
The City of Asylum
Locations (City) (Any)
's comment on 2005-10-27 02:54 PM
Want to give credit where credit is due.
There is a game call Asylum as well. When the world went mad. Interesting, but not really.
Additional ideas...
Perhaps it has been centuries, now the land is farmed by zombie automotons. The only living people in the region are the guards around the city.
Perhaps the entire country is comprised of great cities converted to asylums.
Perhaps there is a gate of somekind in the city where extradimensional things come through and possess the humanfolk.
.....They want to communicate with the sane to save their world, yet their presence causes madness.
.....They would escape the city, but the unique stone of the wall wards them. Talismans of that stone protect you from them.
..... Perhaps they already have escaped the city and have prepared for an invasion. The city is their staging area.
Go to Comment
The City of Asylum
Locations (City) (Any)
's comment on 2005-10-27 02:55 PM
I seem to be on a jag with these orbs.... (See Corvus )
But imagine if a greater orb went mad and took the city with it (given its ability to manipulate people). It could be pushing the regions leader to supporting the poor people of Asylum in its more lucid moments (or asking one of orbs if it would take help the city out).
Even nastier... the orb is telepathic and conducting an experiment on the human minds inside the city (and nearby). It may not be cruel or evil per say, but simply sees the humans as transient lab rats.
Go to Comment
The City of Asylum
Locations (City) (Any)
's comment on 2005-10-27 02:56 PM
Yep. It could be anything. The only people who know for sure are totally mad. I actually like it that nobody knows, it just is. We can put out a number of theories, but we should never say this is right because it would kill the mystery and make it just another puzzle for the players to solve.
Go to Comment
The City of Asylum
Locations (City) (Any)
's comment on 2005-10-27 02:57 PM
Madness can be quite liberating. There is a fine line between madness and genius, and sometimes that line is very, very, blury.
Mad people see the world in different ways, not constrained by binding (or blinding even) paradigns. They can see solutions to problems or innocvations that so called sane people can not seen. It is only those who see the world different who can't relate to those who see the world normally that we lock up (well we should lock up, but now they waunder the streets in and out of halfway houses and medical facilities). Those who can vaugely relate, we pay large sums of money for their interesting ways of looking at the world and the problems that their way of looking at the world can solve.
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Reviews by Faye A. P.
From (Fort Washington, MD)
Sort By: Most Helpful | Newest
the exact color solid- brown i'd been searching for!
This runner was exactly what I'd vision, and had been searching for on the web. I kept getting mixed brown for prices I did not want to pay, or a length that was too short. Eventually, I will get one for another hall, and I will be returning to Floors and Surfaces!
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Air Force preview: It's difficult to win a MW championship
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DOJ investigates cable companies over online video practices
Interview by
Jul 9, 2012
We've all been there: You get home from work, open up your computer to Netflix, press play and.....nothing. The screen reads buffering. Who do you blame? Your internet provider or Netflix?
Watching the L.A. Kings make history
Jun 6, 2012
The Los Angeles Kings could win their first Stanley Cup hockey championship ever tonight. But fans will have to pay a premium to see it whether they watch from the pricey seats at the Staples Center, in a sports bar or at home on cable TV.
Musicians big and small compete for summer concert sales
May 31, 2012
Madonna begins her worldwide summer tour today in Israel. Shows have already sold out around the world. But what if you're not Madonna? How do you make money on tour?
TV Networks sue Dish Network over ad-skipper
May 25, 2012
Satellite TV provider Dish Network has a new digital recorder called AutoHop that cuts out commercials. Fox, NBC, and CBS are now suing to stop it.
Who pays for 3-D movie glasses?
May 23, 2012
The popularity of 3-D movies is growing. So is a battle over who should pay for them -- the studios or the theaters?
Dish Network clashes with TV execs
Interview by
May 17, 2012
The best laid advertising plans could be thwarted by a new service from Dish Network -- the satellite TV operator -- that lets users skip commercials all together.
What makes a movie?
May 16, 2012
These days, a well-established brand can be as valuable as a screenplay. At least that's what the makers of the movie "Battleship" are hoping when the film version of the popular board game opens this weekend.
Biggest suspense from upfronts are the lineups
May 14, 2012
It's upfronts week for the television industry, when networks announce for advertisers which shows are in, on their way in and out. Networking scheduling can be a bit of a "scientific art."
The lowdown on this season's network upfronts
Interview by
May 14, 2012
Variety's Andrew Wallenstein talks about the big trends on television next fall.
ABC, Univision team up for English-language channel
May 8, 2012
The new 24-hour English-language cable channel will target Latinos. But it's an unproven market.


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10-Year-Old Boy Graduates From High School
Description:A 10-year-old boy in Sacramento recently earned his high school diploma. That makes the child, Tanishq Abraham possibly the youngest kid, to graduate from high school. (1:11)
Source: GeoBeats
Tags: GeoBeats , 10 year old diploma , 10 year old graduate , child graduate


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Tuxedo vector background with bow
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Ann Demeulemeester Runway Fall 2012
Ann Demeulemeester Fall 2012
Don't be deterred by Ann Demeulemeester's Fall 2012 tough-girl exterior and Edward Scissorhands-inspired coifs, this collection was chock-full of gorgeously elegant dresses, skirts with expertly deconstructed pleats, and perfectly structured outerwear. Like our antihero Edward Scissorhands, we think this blue- and black-hued lineup shows off a mean-looking exterior at first glance, but the sum of its parts are acutely beautiful with a serious dose of edge and excellent tailoring upon further examination. Be sure to take note of the standout styles: high-necked, floor-sweeping frocks paired with leather opera-length gloves, slick leather pants tucked into slouchy thigh-high moto boots, and biker-inspired jackets featuring slim sleeves and asymmetrical folds. See below for more Ann Demeulemeester details.
- Trends: Deconstructed pleats, strong outerwear, long sweeping dresses, thigh-high boots, and leather pants.
- Colors: Black and midnight blue.
- Key Piece: No Winter wardrobe should be without one of Ann Demeulemeester's utilitarian-chic coats.
- Accessories: Opera-length gloves and thigh-high boots.
- Who Would Wear It: Ladies who prefer strong silhouettes and a dark color palette, like Rooney Mara and Liv Tyler.
- ASAP Rocky is basically a wizard when it comes to clothes — The Cut
- Even accessories have accessories now — Style.com
- Meryl Streep's daughters just landed a cool new fashion gig — HuffPost Style
- Elizabeth Taylor always wore the best jewels — InStyle
- Would you buy an Apple Watch just to make a statement? — Racked
- The shirtdress may be making a comeback this Spring — SheFinds
- Why you should be on board with dresses over pants — Who What Wear
- Kate Spade Saturday is having a crazy-good sale right now — Lucky
- 1 woman weighs in on whether or not we need fashion — Man Repeller


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"Overwhelmingly, businesses are failing to protect themselves by not securing their domain names as trademarks. While the numbers are moving in a positive direction, most businesses on the Internet are at risk that a trademark owner may appear and tell them to 'cease and desist' using that domain name."
Alan Davidson
"To be known and protected by the Lord God from this world of wickedness and insecurity, love Him with all your heart."
Raymond Obeng
"The smart domain-name holders protect themselves in two ways: first, they complete a trademark search before selecting a domain name to ensure availability, and secondly, once they adopt the name, they register it as a trademark to protect themselves from business disruption, ... This way they are covered from all sides."
"unlikely a valid trademark. ... The mark is a weak one as trademarks go."
Denny Chin
"Any franchise is always taking steps to protect their trademarks."
Mark Dawson
"An image . . . is not simply a trademark, a design, a slogan or an easily remembered picture. It is a studiously crafted personality profile of an individual, institution, corporation, product or service."
Daniel J. Boorstin
"When we got back from Athens we found out that they (the COC) had adopted our trademarks."
Jane Roos