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International Journal of Pharmaceutics 476 (2014) 134141
Contents lists available at ScienceDirect
International Journal of Pharmaceutics
journal homepage: www.elsevier.com/locate/ijpharm
Pharmaceutical nanotechnology
Rhamnolipids as emulsifying agents for essential oil formulations:
Antimicrobial effect against Candida albicans and methicillin-resistant
Ester Haba a , Samira Bouhdid c , Noelia Torrego-Solana a , A.M. Marqus a ,
M. Jos Espuny a , M. Jos Garca-Celma b , Angeles Manresa a, *
Unitat de Microbiologia, Facultat de Farmcia, Universitat de Barcelona, Joan XXIII s/n, 08028 Barcelona, Spain
Department of Pharmacy and Pharmaceutical Technology, R+D Associated Unit to CSIC, Faculty of Pharmacy, University of Barcelona, Joan XXIII s/n, 08028
Dpartement de Phytologie, Institut National des Plantes Mdicinales et Aromatiques, Universit Sidi Mohamed Ben Abdellah-Fs, Morocco
A R T I C L E I N F O
A B S T R A C T
Article history:
Received in revised form 16 September 2014
Accepted 26 September 2014
Available online 28 September 2014
This work examines the inuence of essential oil composition on emulsication with rhamnolipids and
their use as therapeutic antimicrobial agents against two opportunistic pathogens, methicillin-resistant
Staphylococcus aureus (MRSA) and Candida albicans. Rhamnolipids, produced by Pseudomonas aeruginosa,
with waste frying oil as the carbon source, were composed of eight rhamnolipid homologues. The
rhamnolipid mixture was used to produce emulsions containing essential oils (EOs) of Melaleuca
alternifolia,Cinnamomum verum, Origanum compactum and Lavandula angustifolia using the titration
method. Ternary phase diagrams were designed to evaluate emulsion stability, which differed depending
on the essential oil. The in vitro antimicrobial activity of the EOs alone and the emulsions was evaluated.
The antimicrobial activity presented by the essential oils alone increased with emulsication. The surface
properties of rhamnolipids contribute to the positive dispersion of EOs and thus increase their availability
and antimicrobial activity against C. albicans and S. aureus. Therefore, rhamnolipid-based emulsions
represent a promising approach to the development of EO delivery systems.
2014 Elsevier B.V. All rights reserved.
Phase behaviour
Methicillin-resistant
Essential oils (EOs) are complex mixtures of volatile organic
molecules extracted from aromatic plants by different methods.
They are oil-like in nature, frequently characterized by a strong
fragrance (Morais et al., 2008) and may contain up to 100 components, mainly terpenes and phenylpropanoids (Hammer and
Carson, 2011). This structural diversity is responsible for the wide
variety of biological activities exhibited by EOs. Several authors
have reported that many EOs, especially those rich in phenols,
aldehydes and alcohols, are effective in inhibiting spoilage and
pathogenic microorganisms (Hammer and Carson, 2011). They
therefore represent a natural alternative to the synthetic
antimicrobials used in the cosmetic, food and pharmaceutical
industries (Lang and Buchbauer, 2012). Their effectiveness means
they can be used in small amounts (Hammer and Carson, 2011).
* Corresponding author. Tel.: +34 93 4024496; fax: +34 93 4024498.
E-mail address: amanresa@ub.edu (A. Manresa).
http://dx.doi.org/10.1016/j.ijpharm.2014.09.039
0378-5173/ 2014 Elsevier B.V. All rights reserved.
One of the most widely studied EO is that derived from
Melaleuca alternifolia, (Myrtaceae), also known as tea tree oil (TTO).
This oil is extracted by steam distillation from the leaves and
terminal branchlets of M. alternifolia and has been used
medicinally for many years. TTO has broad-spectrum in vitro
antimicrobial activity that is mainly attributed to the presence of
terpinen-4-ol and 1,8-cineole, which are both major components
of the oil. TTO is widely used as an antiseptic agent in denture-,
mouth- and hand-wash products. It is also added to topical
formulations for the treatment of cutaneous infections (Carson
et al., 2006; Charles et al., 2013; Sharma et al., 2010).
The EO extracted from Origanum compactum (Lamiaceae), an
endemic species growing in Morocco, has been reported to be
effective in vitro against a wide range of bacteria and fungi,
including pathogenic strains (Bouhdid et al., 2009; Oussalah et al.,
2007). This oil is mainly composed of carvacrol and thymol, which
are phenolic terpene isomers known for their antimicrobial
activity (Ahmad et al., 2011). Cinnamon (Cinnamomum zeylanicum
Blume, syn Cinnamomum verum) belongs to the Lauraceae family.
The EO extracted from cinnamon bark is widely used and has many
applications in perfumery and the food and pharmaceutical
E. Haba et al. / International Journal of Pharmaceutics 476 (2014) 134141
industries (Lee and Balick, 2005). This oil is rich in cinnamaldehyde
and eugenol and exerts several biological functions. Numerous
studies have reported that cinnamon bark oil effectively inhibits
the growth of bacteria and fungi (Bouhdid et al., 2010; Unlu et al.,
2010). Lavender (Lavandula angustifolia), a member of the
Lamiaceae family, produces an EO that is rich in linalyl
acetate and linalool. This oil has been reported to exhibit some
antimicrobial activity against pathogenic microorganisms (de
Rapper et al., 2013; Hammer et al., 1999a).
The industrial application of antimicrobial EOs and their
components is limited by their hydrophobicity, avouring
properties and high volatility (Hyldgaard et al., 2012); it is
therefore important to determine an appropriate formulation for
commercial applications. The pharmaceutical and food industries
have developed several approaches to deliver sensitive antimicrobials (Gudia et al., 2013). Recent studies have reported that some
delivery systems (i.e. emulsion) generally improve the activity of
antimicrobials from herbs and spices and help maintain their
stability (Gaysinsky et al., 2008). An emulsion consists of one liquid
dispersed in another immiscible liquid in the form of droplets, and
is usually stabilized by surfactants that assemble at the interface.
This system may and ensure dispersion of oil in the aqueous phase
where microorganisms are most likely to accumulate. To avoid
possible negative interactions with EO activity, it is important to
choose an appropriate surfactant for the emulsion, since some
surfactants have been found to compromise the antimicrobial
activity of TTO (Hammer et al., 1999b).
Rhamnolipids constitute one of the most important classes of
microbial surfactants and have shown excellent emulsifying
potential with a variety of compounds. They are suitable
candidates for industrial use, since their surface-active properties
remain stable over a broad pH range and when heated (Lovaglio
et al., 2011). Rhamnolipids exert antimicrobial activity against both
fungi and bacteria (Abalos et al., 2001; Haba et al., 2003a; Nitsche
et al., 2005). Their therapeutic applications have been the subject
of studies (Gharaei-Fathabal, 2011; Rodrigues et al., 2006) and
some interesting articles have recently been published (Fracchia
et al., 2012; Gudia et al., 2010).
To our knowledge, no information is available on the use of
rhamnolipids as emulsifying agents in EO formulations intended
for microbial inhibition. The objectives of this study were: (i) to
explore the emulsication behaviour of rhamnolipids with four
commercial EOs (O. compactum, M. alternifolia, C. zeylanicum and L.
angustifolia,) using ternary phase diagrams, and (ii) to assess the in
vitro antimicrobial activity of the emulsions against Staphylococcus
aureus (MRSA) and Candida albicans.
2.1. Microorganisms and growth culture
Pseudomonas aeruginosa 47T2 NCIB 40,044, isolated from an oilcontaminated soil sample, was selected due to its capacity to
produce surface-active rhamnolipids from hydrophobic substrates.
After being grown on TSA (tripticase soy agar, Pronadisa, Barcelona,
Spain), the bacterial strain was maintained at 4  C and also
preserved in cryobilles (AES Chemunex S.A., Terrassa, Spain) at
20  C.
Experiments were carried out in 2-l bafed Erlenmeyer asks
containing 400 ml of medium with the following composition (g/l):
NaNO3 5, KH2PO4 2.0, K2HPO4 1.0, KCl 0.1, MgSO47H2O 0.5, CaCl2
0.01, FeSO47H2O 0.012 and yeast extract 0.01. We added 0.05 ml of a
trace element solution containing (g/l): H3BO3 0.26, CuSO45H2O
0.5, MnSO4H2O 0.5, MoNa2O42H2O 0.06 and ZnSO47H2O 0.7 to
this medium. Finally, 40 g/l of olive/sunower (50:50 v/v) waste
frying oil was used as a carbon source, containing oleic acid
(50.29%) and linoleic acid (34.23%) as the major components, and
stearic acid (7.70%) and palmitic acid (7.77%) in smaller quantities.
The medium components were sterilized separately at 120  C,
1 atm for 20 min. The initial pH of the medium was adjusted to 7.2.
A 2% (v/v) cell suspension in sterile saline (0.9% NaCl) of an
overnight culture on TSA (Pronadisa, Barcelona, Spain) was used as
the inoculum. Cultures were incubated at 30  C for 96 h on a
reciprocal rotary shaker at 150 rpm.
2.2. Rhamnolipid production and characterization
Microbial growth was calculated by measuring the protein
content of the cultures in accordance with the method described
by Lowry et al. (1951). Total rhamnolipid (RL) production was
measured as rhamnose by a specic colorimetric method
(Chandrasekaran and Bemiller, 1980). The RL content was
calculated by multiplying the rhamnose concentration by a factor
of 3.0, which represents the rhamnolipid/rhamnose calculated
using the puried product.
Rhamnolipids were recovered and puried. Cells were removed
from the culture by centrifugation (12,000  g) for 30 min.
Purication was carried out by adsorption chromatography
(Torrego-Solana et al., 2014).
Liquid chromatography-mass spectrometry (LCMS) was
performed in order to analyse the composition of the RL mixture,
and a 10-mg portion was resuspended in 1 ml of methanol and
analysed by LCMS. Rhamnolipid mixtures were separated and
identied by LCMS using a Waters 2690 separation module
(Waters, Milford, MA, USA). Samples were injected (10 ml) into a
C18 Spherisorb ODS2 150  4.6-mm column (Teknokroma, Sant
Cugat, Spain). The LC ow rate was 1 ml min 1. An acetonitrile
water gradient was used, starting with 30% acetonitrile for 2 min,
followed by a ramp of 30100% acetonitrile for 30 min, before
being left to stand for 5 min and then returned to the initial
conditions. Post-column addition of acetone at 200 ml/min was
performed using a Phoenix 20 syringe pump (Carlo Erba, Rodano,
Italy), since we observed a rise in sensitivity to rhamnolipids. The
LC efuent and acetone were mixed in a tee valve (Valco) and split
(1/50) before being introduced into the mass spectrometer. MS
was performed with a single quadrupole mass spectrometer, VG
Platform II (Micromass, Manchester, UK), equipped with a
pneumatically assisted electrospray (ES) source. Negative ion
mode was used (N-ES). Full scan data were obtained by scanning
from m/z 100 to 750 in centroid mode, using a scan duration of
2.0 s and an inter-scan time of 0.2 s. The working conditions for NES were as follows: dry nitrogen was heated to 80  C and
introduced into the capillary region at a ow rate of 400 l/h.
The capillary was held at a potential of 3.5 kV and the extraction
voltage was held at
80 V. Rhamnolipid homologues were
quantied from the molecular proportion of each of the
pseudomolecular ions calculated by LCMS (Haba et al., 2003b).
The proportion (%) of each component was calculated from the
areas obtained for each molecule of RL by LCMS.
Equilibrium surface tension (g ST) and critical micelle concentration (CMC) were measured at 25  C by the DuNoy ring method
with a Krss K9 tensiometer (Hamburg, Germany). The instrument
was calibrated against Milli-Q ultrapure distilled water (Millipore,
Billerica, MA, USA). Aqueous solutions with various concentrations
of biosurfactants (2005 mg/l) were obtained by successively
dilutions prepared by weight in Millipore ultrapure water for CMC
determination. To reach equilibrium, all sample solutions were
aged in appropriate cells at room temperature (25  C). The
platinum plate and all glassware used were cleaned in chromic
mixture. CMC was calculated from surface tension plot values
versus log surfactant concentration after reaching equilibrium
at 25  C.
2.3. Essential oils
The four EOs (supplied by Pranarm, Ghislenghien, Belgium)
used for this study were oregano essential oil (OEO) extracted from
O. compactum ower heads from Morocco, cinnamon essential oil
(CEO) extracted from the bark of C. zeylanicum from Sri Lanka, tea
tree oil (TTO) extracted from the leaves of M. alternifolia from
Australia, and lavender essential oil (LEO) extracted from ower
heads of L. angustifolia from France. Oils were commercialized and
chemotyped by Pranarm International (Ghislenghien, Belgium).
According to the supplier, the major components of the oils were:
carvacrol (37.77%), thymol (19.79%) and g-terpinene (17.01%) for
OEO; E-cinnamaldehyde (77.31%) for CEO; terpinen-4-ol (39.39%)
and g-terpinene (20.76%) for TTO; and linalool (30.60%) and linalyl
acetate (34.09%) for LEO). The GC and the physical characteristics of
the EOs are included (Supplementary information). All EOs were
used as supplied.
2.4. Determination of emulsifying properties of rhamnolipids in water/
oil mixtures
The emulsifying properties of puried RL were determined
following the titration method and represented in ternary phase
diagrams (Sadurn et al., 2005). Non-equilibrium phase diagrams
were performed at 25  C by stepwise addition of water to the RL
and oil mixture. Emulsions were prepared by weighing the
components up to 1 g as follows: the required concentrations of
RL were weighed in glass tubes, mixed with the required amount of
the hydrophobic component and vortexed until complete dissolution of the biosurfactant was achieved. The appropriate amount of
deionized water was then added slowly and the tube was tightly
stoppered and mixed vigorously for 1 min. The shaking time and
vortex speed were kept constant. Finally, the mixtures were kept in
a thermostatic bath at a constant temperature of 25  C for 24 h. The
emulsion boundaries were determined by visual inspection. The
samples were examined to determine if a clear single phase formed
immediately or phase separation occurred, or an opaque mixture
appeared, which would indicate the coexistence of two or more
phases. The optical anisotropic aspect was detected with crossed
polarizers.
The physical aspect of the mixtures was plotted on a ternary
phase diagram with three vertexes representing water, RL and oil
(W/RL/EO). The emulsion region was represented. The selected
hydrophobic components assayed in this study were the four EOs
in the range of 0.590%. The maximum amount of biosurfactant
was 20% (from 0.1% to 20%). The dots of emulsions on the phase
diagram were coded and the percentage of each component given
(%W/%RL/%EO); the starting points were: (75-20-5); (66-20-14);
(50-20-30); (34-20-46); (0-20-80); (0-10-90). The results obtained
with the different oils studied were compared and the emulsions
were also characterized by optical microscopy (Leica DM IL LED,
Leica Microsystems Barcelona, Spain). Images were acquired with a
Leica EC3 digital camera and the software Image.
2.5. Minimum inhibitory concentration (MIC) of essential oils and
First the antimicrobial activity, based on MICs, of the EOs and
rhamnolipids was determined; for C. albicans it was ATCC 10231
and for methicillin-resistant S. aureus (MRSA) it was ATCC
MICs were determined using the broth microdilution assay
(Bouhdid et al., 2009): rst, 50 ml of Mueller Hinton Broth (MHB,
Oxoid, Basingstoke, UK) or Sabouraud-dextrose broth (SDB, Oxoid,
Basingstoke, UK) supplemented with bacteriological agar (0.15% w/
v) was distributed from the second to the 12th well of a 96-well
polypropylene microtitre plate (Costar, Corning Incorporated,
Corning, NY, USA). A dilution of the EO was prepared to a nal
concentration of 4% in the corresponding medium. Then, 100 ml of
these suspensions were added to the rst test well of each
microtitre line, and then 50 ml of scalar dilution was transferred
from the second to the 11th well. The 12th well was considered as
the growth control, because no EO was added. We then added 50 ml
of a microbial suspension to each well at a nal concentration of
approximately 105106CFU/ml. The nal concentration of EO was
between 2% and 0.0019% (v/v). Plates were incubated at 37  C for
18 h. After incubation, 5 ml of resazurin (0.01% w/v) was added to
each well to assess active microbial growth. After further
incubation at 37  C for 2 h, the MIC was determined as the lowest
EO concentration that prevented a change in resazurin colour.
Active microbial growth was detected by reduction of blue dye
resazurin to pink resorun. A control was carried out to ensure that
the EO did not cause a colour change in the resazurin at the
concentrations tested. Experiments were performed in triplicate
and modal values were selected. The same method was followed
for the RL, although nal concentrations of rhamnolipids ranged
from 256 mg/ml to 0.0244 mg/ml. Vancomycin hydrochloride
(SigmaAldrich, Saint Louis, USA)) and amphotericin B (Sigma
Aldrich, Germany) were used as the control.
2.6. Antimicrobial activity of the emulsions by agar-well diffusion
The antimicrobial activity of the selected emulsions and
individual components was assessed by agar-well diffusion. After
solidication of a basal layer (20 ml) of sterile Mueller-Hinton agar or
Sabouraud dextrose agar (Oxoid, Basingstoke, UK) in Petri dishes,
sterile 8-mm diameter cylinders were deposited on top. Then, 6 ml of
LB medium (Oxoid, Basingstoke, UK) containing 0.8% agar was
inoculated with a fresh culture of the microbial strain (nal
concentration 106 CFU/ml) and poured over the surface of the
medium. After solidication, the cylinders were pulled out and the
wells were lled with 50 ml of the chosen emulsions. After
incubation at 37  C for 24 h, all plates were examined for any region
of growth inhibition, and the diameter (mm) of these regions was
measured (Bouhdid et al., 2008) All tests were performed in
triplicate. The emulsions were prepared as described previously and
placed into the wells immediately after preparation. In addition, RL
and EOs were tested individually. The volumes applied to the wells
were equivalent to the concentration in the emulsions tested.
3.1. Rhamnolipid characterization
In order to reduce the cost of rhamnolipid production, prolong
the life of the materials and therefore minimize the environmental
impact, edible oil waste was used for microbial conversion to
produce biosurfactants (Abalos et al., 2004; Benincasa et al., 2004;
Haba et al., 2000; Mercad et al., 1993).
After RL purication, a sticky, semi-solid, brown-coloured oil
product with 98% purity was obtained. The composition of the
surfactant produced by P. aeruginosa 47T2 was determined by LC
MSES to be a mixture of eight homologues (RL8). As shown in
Table 1 most of the accumulated rhamnolipid (60.4%) were monorhamnolipids: R1-C8-C10; R1-C10-C10; R1-C10-C12; R1-C10-C12:1 Being
the major component R1-C10-C10 (39.14%). The 39.6% of the mixture
were di-rhamnolipids: R2-C8-C10; R2-C10-C10; R2-C10-C12; R2-C10C12:1; being the main componentnt R2-C10-C10 (19.18%). Accumulation started soon after incubation and lasted until the end of the
process (96 h), whereas the minor homologues accumulated after
growth ceased.
Rhamnolipid homologues RL8 produced by P. aeruginosa 47T2. Relative abundances (%) were calculated from the pseudomolecular ion peak areas.
Rhamnolipid
homologues
Pseudo-molecular ion (m/z)
Relative abundance (%)
Fragments (m/z)
R-C10-C10
R2-C10-C10
R2-C10-C12/R2-C12-C10
R-C10-C12/R-C12-C10
R-C10-C12:1/R-C12:1-C10
R2-C10-C12:1
R2-C8-C10
R-C10-C8/R-C8-C10
333,169, 119, 103
479, 169,163
361, 333,169, 163, 119, 103
305, 169, 163, 141, 119,103
Rhamnolipids accumulated in the culture medium as a mixture
of different homologues whose nal composition appeared to
depend on the bacterial strain and substrate composition.
Although rhamnolipid properties are known to depend on the
distribution of their homologues, little is known about the
contribution of each individual homologue to the surface
properties of rhamnolipid mixtures. The new surfactant (RL8)
reduced the surface tension to 32.21 mN/m and the critical micelle
concentration (CMC) observed was 105 mg/l. The CMC value for
RL8 appeared to be slightly lower than that observed with a
rhamnolipid mixture of 11 homologues (108 mg/l) produced by the
same strain (Haba et al., 2003a). This could be due to the smaller
amount of unsaturated components of the surfactant produced in
the present study (9.88%) compared to the 13.87% reported
previously (Haba et al., 2003a). The effect of the presence of
unsaturated compounds was also observed in the case of strain LBI,
which contained up to 31% unsaturated carbon with a CMC of
120 mg/l (Benincasa et al., 2004), and also for the AT10 strain,
which contained up to 43.2% unsaturated compounds with a CMC
of 150 mg/l (Abalos et al., 2001). However, despite differences in
the composition of individual homologues and CMC values, surface
tension values (2732 mN/m) and HLB values (710) were in the
same range of hydrophobicity, and therefore the same range of
emulsication behaviour (Attwood and Florence, 1983); in the case
of RL8, an HLB of 8.13 was calculated, which favours O/W
microemulsions.
3.2. Emulsifying properties of RL47T2 in water/oil mixtures
Biosurfactants have been shown to have valuable biological
properties and several pharmaceutical (Gharaei-Fathabal, 2011)
and biomedical applications (Rodrigues et al., 2006; Stipcevic et al.,
2006). They have been reported to have antimicrobial, antiadhesive and other biological applications (Cameotra and Makkar,
2004; Das et al., 2009; Fracchia et al., 2012; Gharaei-Fathabal,
2011; Gudia et al., 2010; Haba et al., 2003b; Kitamoto et al., 1993).
The trend for using natural emulsifying agents in pharmaceutical
formulations favours the use of biosurfactants as an alternative to
their chemical counterparts, and these show promise for the future
(Gudia et al., 2013). By exploiting the excellent emulsication
behaviour of rhamnolipids in an attempt to broaden their range of
applications in therapeutic EO formulations, ternary systems were
plotted to study the emulsifying capacity of RL8 and one of the EOs
of O. compactum (OEO), C. zeylanicum (CEO), M. alternifolia (TTO)
and L. angustifolia (LEO) and water (Fig. 1ad). The ternary phase
diagrams provide in-depth knowledge about what a system may
offer in terms of the phase behaviour, depending on the relative
proportions of its components.
Despite the fact that the phase behaviour of water/surfactant/
oil systems at constant temperatures has been studied extensively
for pharmaceutical and cosmetic formulations (Attwood and
Florence, 1983), and for agricultural and food applications, the
literature provides little information on phase diagrams with
biosurfactants (Abalos et al., 2004; Kitamoto et al., 2009; Marqus
et al., 2009; Torrego-Solana et al., 2014).
OEO, TTO and LEO have a lower density than water; they are
immiscible in water and phase separation was apparent. Although
TTO is slightly soluble in water, some turbidity was observed in the
composition studied due to the low solubility (Morais et al., 2008).
In terms of the miscibility of RL8 with the EOs, RL8 was found to be
soluble in CEO and TTO. While rhamnolipids are soluble in water,
the lipophilic nature of EOs promotes phase separation. Since the
role of biosurfactants as emulsifying agents is to ensure the
dispersion of EOs in water, and low concentrations are needed, no
more than 20% of RL8 was added. Destabilization of emulsions
observed after few minutes of preparation in emulsions with TTO
and OEO could be attributed to Ostwald ripening. In oil-in-water
emulsions, the kinetics of Ostwald ripening depends on the
diffusion of the oil molecules across the aqueous phase separating
droplets (Suriyarak and Weiss, 2014). This destabilization process
is of particular importance for emulsions containing lipids with
appreciable water solubility (e.g. avours and essential oils).
Emulsions of essential oils, which consist of various terpenes, can
show distinct Ostwald ripening kinetics.
Oregano oil is dark brown in colour and, together with RL8,
produces the brownish colouration observed in some emulsions.
Examination of the ternary phase diagrams showed that, in the
OEO system (W/RL/OEO), two main areas with two different
behaviours were distinguished (Fig. 1a). In zone A, the emulsions
contained up to 40% OEO, 120% RL and 5095% water. Visual
inspection revealed thick, cream-coloured emulsions, most of
which presented creaming phenomena.With a low water concentration and 520% RL8, a different area (zone B), composed of more
than 50% emulsion and water separation at the bottom, was
observed; these emulsions were thick and dark brown in colour.
The emulsions were consistent in both areas and when observed
under optical microscopy multiple emulsions were visible (Fig. 2a).
This phenomenon has been described previously in emulsions of
rhamnolipids from strain AT10 and Casablanca crude oil, which is
also a complex substrate with more than 26% aromatic compounds
(Abalos et al., 2004).
TTO formulations of a monoolein/water system as a carrier for
terpinen-4-ol (Caboi et al., 2002) and more recently, the
development of hydrogel-thickened nanoemulsions with vitamin
A palmitate (Oliveira et al., 2011) have been reported, no other
information on the behaviour of TTO in biologically active
formulations was found. In the TTO system (W/RL/TTO), zone A
was smaller than in the OEO system (Fig. 1b). This zone extended
from 55% water to the water vertex, with up to 30% of the oil being
solubilized. In contrast to the OEO system, the maximum amount
of surfactant used was 9%. The excess of water made the emulsions
appear light and milky. Above this area, when TTO was in the range
25% and RL was over 9%, gels rather than emulsions were
observed. Birefringence was observed under polarized light for
some compositions with a water concentration ranging from 73%
to 93% and 5% TTO, that could be attributed to the formation of
Fig. 1. The pseudo-phase diagrams show the emulsion areas for the OEO (a), TTO (b), CEO (c) and LEO (d) systems. White dots indicate monophasic compositions. The
intersection (black dots) between the dotted lines show the emulsion used to study the antimicrobial activity of the emulsions.
anisotropic structures, such as lamellar or hexagonal liquid
crystals. These results differ from those reported to obtain stable
emulsions with TTO/TS weight ratios (%) 15/10, 10/5, 15/5 and 20/
10, points outside the best-area boundaries found in this work. This
might be due to the surfactant used, a mixture of nonionic
surfactants (ethers), whereas rhamnolipids belong to the carboxylic esters in the anionic group (Morais et al., 2008).
In the case of the CEO system (W/RL/CEO), the emulsions were
not formed in the same way as the OEO or TTO systems. As shown
in Fig. 1c, despite the fact that RL8 was soluble in CEO, phase
separation was apparent in all compositions studied. Even at high
water concentrations a dark phase appeared at the bottom in
equilibrium with a uid emulsion. This behaviour could be
explained by the high percentage of E-cinnamaldehyde (77.31%),
which is barely soluble in water, as well as the low proportion of
oxygenated monoterpenes such as 1,8-cineole (0.16%), linalool
(2.8%) and a-terpineol (0.27%) in the oil (Edris and Malone, 2011).
Observation of the bottom phase under optical microscopy showed
a compact and homogeneous emulsion (Fig. 2b). In the LEO system
(W/RL/LEO), the best emulsions were observed when the EO
ranged between 0.7% and 1.6% and RL between 3% and 7.5%, and the
weight ratio for water was over 91% (Fig. 1d, zone A). Milky
emulsions with a thin creamy lm formed in the aqueous region. A
multiphase area (zone B) near the oil vertex formed from the EO/RL
axis to near 50% water.
EOs are multi-component systems, and therefore their solubilization and consequent emulsication result from the interaction
of all minor and major constituents, with each other and with the
surfactant at the interface layer (Edris and Malone, 2011).
Comparing the four systems studied, OEO produced the thickest
Fig. 2. Emulsions observed under optical microscopy. Emulsion A, formed from OEO, is an exemple of multiple emulsion W/OE/W. Emulsion B, would be an example of
emulsion formed from CEO, TTO or LEO.
Minimal inhibitory concentration MIC % (v/v). The values presented belong to each
essential oil against C. albicans and MRSA S. aureus. OEO and CEO presented same
antimicrobial activity against both microorganisms.
C. albicans
MRSA S. aureus
Oreganum oil (OEO)
Cinnamomum oil (CEO)
Tea tree oil (TTO)
Lavander oil (LEO)
emulsions, while TTO and LEO produced light, milky emulsions
with less than 9% RL. For the OEO, TTO and LEO systems, the best
emulsions were formed when water was over 50% and EO content
was below 40%.
3.3. Antimicrobial effect
3.3.1. Minimum inhibitory concentration (MIC)
The EOs used were rst examined for their antimicrobial effect.
Specically, their minimum inhibitory concentrations (MIC) were
determined against two pathogens: C. albicans and methicillinresistant S. aureus (MRSA). The MICs were assayed in liquid
medium and a small amount (0.15%) of agar was added to prevent
phase separation (Mann and Markham, 1998). As shown in Table 2,
CEO and OEO showed the highest antimicrobial activity against
MRSA, with an MIC value of 0.125% (v/v), whereas the activity
against C. albicans revealed MIC values of 0.0156% and 0.008% (v/v)
for CEO and OEO, respectively. Regarding the other oils, TTO and
LEO inhibited C. albicans at an MIC value of 0.5% (v/v) and MRSA at
2% (v/v). These results are consistent with those obtained over
44 resistant clinical isolates and against signicant pathogenic
bacterial and fungal isolates from the oral cavity and skin (Warnke
et al., 2009; Warnke et al., 2013). The MICs of RL8 were higher than
256 mg ml 1 against both of the microorganisms tested. Vancomycin hydrochloride inhibited MRSA at an MIC value of 0.125 mg ml 1,
while C. albicans was inhibited by amphotericin B at 0.065 mg ml 1.
3.3.2. Antimicrobial effect of the emulsions
In order to prevent disaggregation, the antimicrobial activity of
EO emulsions was studied by means of the diffusion test, rather
than the dilution method. To our knowledge, this is the rst time
that rhamnolipids, low-molecular-weight biosurfactants, have
been used as an emulsifying agent in an EO formulation.
The emulsions tested all had a high EO content that was within
the therapeutic range. The following emulsions were selected
(% W/RL8/EO): OEO, 72.2/11.1/16.7; TTO, 71.8/2.8/25.3; CEO, 80.9/
1.9/17.1; and LEO, 78.7/8.5/12.8 (Fig. 1ad). Single components,
RL8 and EOs, were also tested as controls due to their inhibitory
effects at the concentrations used in the chosen emulsions
(Table 3). As shown, RL8 showed some inhibition (9.0 mm) at
2.8% and 8.5% (w/v) (Table 3) for both strains. This effect reached
10.0 mm when the concentration was increased to 11.1% (w/v).
The inhibition zone measured for EOs at the concentration used
in the emulsion ranged from 21.3 mm to 37.3 mm for C. albicans
and from 11.0 mm to 24.6 mm for MRSA. No inhibition effect was
found with LEO. OEO was the most effective against C. albicans
(OEO > CEO > TTO), while the order of effectiveness against MRSA
was CEO > OEO > TTO (Table 3). In terms of emulsion systems, the
CEO emulsion was the most effective against both microorganisms,
with an inhibition zone of 42.8 mm and 24.2 mm for C. albicans and
MRSA, respectively, followed by the OEO and TTO emulsions. The
least effective emulsion was LEO (10.0 mm), i.e. CEOe > OEOe >
TTOe > LEOe.
According to Rota et al. (2008), the antimicrobial activity of EOs
can be classied into three levels, depending on the inhibition zone
diameter: weak activity (inhibition zone 12 mm), moderate
activity (12 mm < inhibition zone < 20 mm) and strong activity
(inhibition zone 20 mm) (Rota et al., 2008). Thus, RL8 emulsions
with OEO and CEO have strong activity against C. albicans and S.
aureus (MRSA), while the TTO emulsion has strong activity against
C. albicans and moderate activity against S. aureus (MRSA). The LEO
emulsion showed weak activity in both cases.
From the results obtained, we can conclude that the EOs used in
this study (OEO, TTO, CEO and LEO) could be used topically, since
the oil concentrations fall within the safe interval. Moreover, with
the inclusion of RL in category IV of the EPA classication, which
includes non-irritant products (Haba et al., 2003b), they could
yield some very useful but simple formulations.
The nature of the solubilizing agent is one of the physicochemical parameters that may affect the antimicrobial effect of EOs.
High-molecular-weight surfactants such as Tween 80 or Tween 20,
which are used in many in vitro antimicrobial assays, may
compromise the antimicrobial effect of EOs. This effect is due to
the possible encapsulation of oil in surfactant micelles that
prevents the agent from interacting with microorganisms (Hammer and Carson, 2011; Hammer et al., 1999b; Lang and Buchbauer,
2012). In the present study, we found that rhamnolipids (lowmolecular-weight surfactants) enhance the antimicrobial effect of
the EOs tested. In fact, RL micelles may transport EOs through the
aqueous phase to the surface of bacterial cells and thus improve
contact between the bacterial membrane and EO components. It is
generally accepted that the treatment of microorganisms by EOs
results in the impairment of membrane integrity and function
(Hammer and Carson, 2011). Thus, the hydrophobicity of EOs and
their components allows them to diffuse through the cell wall of
gram-positive bacteria and fungi and the outer membrane of gramnegative bacteria. They enter the phospholipid bilayer of the
bacterial cell membrane, which results in the alteration of the
physical properties of the membrane. Terpene accumulation leads
to the swelling of the lipid bilayer and hence a rise in membrane
uidity (Sikkema et al., 1994). Membrane damage is veried by a
collapsed membrane potential, inhibited respiration, potassium
leakage, loss of intracellular material, including DNA and proteins,
and ultrastructural alterations (Bouhdid et al., 2010; Carson et al.,
2002; Chami et al., 2005).
In recent years, emulsions and nanoemulsions have been
studied as carrier systems for hydrophobic antimicrobials such as
EO components (Dons et al., 2011; Sugumar et al., 2014 Wu et al.,
2014). The effectiveness of emulsions as hydrophobic drug carriers
seems to be affected by the emulsion's surfactant nature and
Growth inhibition zone for the neat oil (EO), RL and emulsion (W/RL/EO). Inhibition zone diameters (mm) produced around the wells by adding 50 ml of the emulsion and the
amount of RL and EO equivalent to the one used in the emulsion. Values are means of three measurements. NI: indicates no inhibition.
EO Emulsion
W/RL/EO
OEO: 72.2/11.1/16.7
TTO: 71.8/2.8/25.3
CEO: 80.9/1.9/17.1
LEO: 78.7/8.5/12.8
C.albicans ATCC 10,231
S. aureus ATCC 43300 (MRSA)
10.0  0.0
droplet size (Dons et al., 2012; Terjung et al., 2012). In fact,
complex interactions between microorganisms and the emulsion
may take place. In some cases, reducing the size of the micelles
improves their interaction with the target microorganisms. In
other cases, when antimicrobials are strongly bound to the droplet
interfaces and the droplets do not interact directly with the
microbial cells, antimicrobials may no longer be released from the
micelle and therefore fail to inhibit microorganisms (Weiss et al.,
These results led us to suggest that formulations of the assayed
EOs could be used in the treatment of candidiasis and MRSA
infections as alternatives to well-established drugs. Furthermore,
we demonstrated that rhamnolipids are highly suitable surfactants
in the context of EO emulsication. Specically, their surface
properties promote the dispersion of EOs, thereby increasing their
availability and antimicrobial activity against C. albicans and MRSA.
Rhamnolipid-based emulsions present a promising avenue in the
development of EO delivery systems. Our study provides preliminary results for further research into the incorporation of
rhamnolipids in different EO-based formulations designed to
enhance the antimicrobial effect of EOs.
The nancial support of the Comisin Interministerial de
Ciencia y Tecnologia (CICYT), project CTQ 2010-21183-C02-01/PPQ
of the Ministerio de Educacin y Ciencia, Spain, and the Comissi
Interdepartamental de Recerca i Tecnologia (CIRIT), Generalitat de
Catalunya project 2014SGR534, is gratefully acknowledged.
Appendix A. Supplementary data
Supplementary data associated with this article can be
found, in the online version, at http://dx.doi.org/10.1016/j.
ijpharm.2014.09.039.
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Emulsione
Membrana cellulare
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The ICCSD staff giving campaign kicks off on December 3.
The 2018-2019 ICCSD staff giving campaign will once again benefit two organizations that directly impact our students by enriching their experience in the classroom and by breaking down barriers to education outside the classroom. ICCSD staff are asked to make a difference in our district and community by supporting the Iowa City Community School District Foundation and the United Way of Johnson and Washington Counties by giving at least $1 per month to each organization.
Staff donation forms for the campaign are due by December 14.
New this year! Donors giving at least a minimum of $60.00 to the ICCSD Foundation are eligible for daily prize giveaways. Turn in your form early to be eligible for the drawing each day!


text:
- OUR EVOLUTION
- RUBICON CONSUMER HEALTHCARE
- RUBICON ACADEMY
- MANUFACTURING
VENTURE WITH US
Rubicon Research and General Atlantic Announce Strategic Partnership
Thane/Mumbai(India), 8th Apr 2019
Rubicon Research Pvt Ltd ("Rubicon"), a global leader in innovative pharmaceutical product development, announced today that General Atlantic has become a significant shareholder in the company. General Atlantic, a leading global growth equity firm, is committing up to $100 million of primary capital and has acquired the stake held by Rubicon's existing private equity shareholders. All other existing shareholders continue to maintain their shareholding in the company.
Founded in 2001, Rubicon has established itself as a well-recognized outsourcing partner to the global healthcare and pharmaceutical industry. Over the past few years, Rubicon has expanded its development expertise with a commercial focus, leading to multiple product approvals and launches. With this investment from General Atlantic, Rubicon intends to focus its efforts on bringing to market innovative medicines to solve fundamental patient needs through the use of its proprietary drug delivery technologies, as well as increase its manufacturing footprint across the world. Further, General Atlantic is partnering with Rubicon to create an innovation platform through both strategic organic and inorganic growth initiatives. Rubicon is launching a "Grow Together" initiative through which Rubicon will back, with both capital and strategic support, seed and early-stage ventures founded by entrepreneurs and innovators in the traditional pharmaceutical domain as well as allied domains involving the application of emerging technologies like AI, continuous manufacturing and big data in the pharmaceutical industry, to achieve Rubicon's key goals of Innovation, Quality and Care. Proposals received by Rubicon from such entrepreneurs and innovators from across the globe, who are looking for financing and partnerships, will be confidentially reviewed by the CEO's office to evaluate their fit into Rubicon's stringent science criteria.
"We welcome this partnership with General Atlantic and believe that with their growth focus, capital resources, strategic outlook and global connectivity, Rubicon will be able to accelerate its growth through organic and inorganic efforts. With initiatives like "Grow Together", Rubicon intends to bring to market innovative technologies and medicines to improve the quality of patient's lives", said Mrs. Pratibha Pilgaonkar, Founder and Chief Executive Officer of Rubicon.
Sandeep Naik, Managing Director and Head of India & Asia-Pacific, of General Atlantic, and Shantanu Rastogi, Managing Director of General Atlantic, focusing on investments across the firm's core sectors of Financial Services, Healthcare, Consumer and Technology in India & the Asia-Pacific region, will join the board of Rubicon. "We have been extremely impressed by the Rubicon team and the quality of the company's R&D. Rubicon's innovative technology platforms are a real differentiator and position the company to sustain its strong growth in the complex and specialty generics market", said Sandeep Naik. General Atlantic will work with Rubicon on a number of strategic initiatives including inorganic growth and geographic expansion of the company's existing footprint. "We are excited to partner with Mrs. Pilgaonkar and the entire Rubicon team to build Rubicon into an innovation-led leader in the global pharmaceutical market", added Shantanu Rastogi.
Torreya Partners, a boutique healthcare investment bank, was the sole financial advisor for Rubicon on this transaction.
About Rubicon Research Pvt Ltd
Rubicon is a responsible partner to global healthcare innovators that design and deliver quality products for making patients' lives healthy and happy. Rubicon also develops and commercializes its own value added formulations in the global market. Rubicon currently has several commercial products in the global market and a pipeline of over 60 products in development or under review. Rubicon holds more than 50 issued patents in drug delivery technologies and with its new R&D centre can develop pharmaceutical products across dosage forms. The manufacturing facility located in Ambernath (India) has global regulatory approvals for commercial supply of oral solid dosage forms. Rubicon also has a marketing office based in New Jersey, USA.
For more information on the "Grow Together", please visit : www.rubicon.co.in/venture
www.rubicon.co.in
About General Atlantic
General Atlantic is a leading global growth equity firm providing capital and strategic support for growth companies. Established in 1980, General Atlantic combines a collaborative global approach, sector specific expertise, a long-term investment horizon and a deep understanding of growth drivers to partner with great entrepreneurs and management teams to build exceptional businesses worldwide. General Atlantic has more than 150 investment professionals based in New York, Amsterdam, Beijing, Greenwich, Hong Kong, Jakarta, London, Mexico City, Mumbai, Munich, Palo Alto, São Paulo, Shanghai, and Singapore.
For more information on General Atlantic, please visit the website: www.generalatlantic.com
HOME KNOW US PRODUCTS PRESS RELEASES CONTACT US
Corporate HQ and R&D Center
Rubicon Research Private Limited
MedOne House, Plot No. B-75, Road No. 33, Wagle Estate,
Thane (W) 400604,
Contact: +91 22 61414000
Plot no. K 30/4 & 30/5,
Additional MIDC, Ambernath,
District Thane 421 506,
Contact: +91 251 2628100
Suite 605, Princeton Office Center II,
666 Plainsboro Road, Plainsboro,
Canada R&D Center
Rubicon Research Canada Limited
255 Spinnaker Way,
Unit 6, Concord,
Ontario, L4K 4J1,
Rubicon Consumer Healthcare Private Limited
Rubicon Academy LLP
Copyright © 2021. Rubicon Research Pvt. Ltd. All rights reserved

redpajama_set_name: RedPajamaCommonCrawl


EU legislation on air quality has been around since the eighties, but the early writings were more of a recommendatory nature, with little legal space to act on countries that were in non-compliance. In the air quality framework directive from 1996 (1996/62/EC) the basic conditions for legally binding air quality standards were introduced, but it was not until 2005 that air quality standards for particulate matter (PM10) came into full effect and any legal claims could be made. For nitrogen dioxide (NO2) the standard entered into effect in 2010 (with the option for countries to request a time extension until 2015). Limits for PM2.5 will not be binding until 2015.
In 21 of the EU member states at least one of their air quality zones exceeded the 24-hour limit for PM10 in 2009. The regular routine is for the European Commission to report countries that fail to comply with the directive to the European Court of Justice (ECJ). So far, only two countries, Sweden and Slovenia, have been condemned for exceeding the PM10 limits in 2005, 2006 and 2007, but not given any penalties. Cyprus, Italy, Portugal, Spain, France and Belgium have all been referred to the ECJ but are still awaiting court decisions. Another group of countries have received warnings.
However not all of the 20 per cent of the EU urban population that lives in areas where PM levels are too high have the patience to wait for years for the EU bureaucracy to run its course. By suing a city, using the national legalisation that follows the transposition of EU legislation, or if needed going all the way to the ECJ, civil society has the possibility to take action.
Munich was one of the first cases of this type. According to German law, environmental organisations cannot appeal against environmental legislation – this can only be done by affected citizens. A Mr Dieter Janecek, who lives near the central ring road in Munich, volunteered to be this individual. Close to his home was one of the municipality's monitoring stations. In 2005 and 2006, high levels of PM10 were recorded there, for more than the 35 days allowed in the directive.
Mr Janecek demanded that the city should adopt an action plan to deal with the air quality problems in his district. The application was dismissed in the court of first instance. He continued to a higher administrative court, which stated that a citizen has the right to require an action plan, but cannot insist on measures that would guarantee compliance in the short term.
Both Mr Janecek and his counterpart, the state of Bavaria, appealed against this judgement to the Federal Administrative Court, which stated that national law alone cannot entitle an individual to have an action plan drawn up. The court however directed a question to the European Court of Justice (ECJ), asking whether conditions were different under Community law. In 2008 the ECJ confirmed that individuals directly concerned can require local authorities to develop short-term action plans when there is a risk that alert thresholds or limit values are exceeded.
With the outcome of the ECJ behind them, German citizens have pursued cases against several other cities that exceed air quality standards, thus forcing them to develop or renew action plans.
A similar case occurred in Sweden, where the Swedish Society for Nature Conservation (SSNC) has pursued a court case on behalf of five members living near one of central Stockholm's main streets, known for having the worst air quality in the country. Due to the heavy transit traffic and poor air circulation the air quality limits for PM10 and NO2 are exceeded every year.
In January 2008, SSNC handed a complaint to the City of Stockholm, with a demand to act on the high levels of air pollution. In April 2009, the municipality rejected the complaint on the grounds that further investigations were needed. A month later, SSNC lodged an appeal to the County of Stockholm. In June 2011 the County Administrative Board set aside the City of Stockholm's decision to reject complaints without further action regarding the levels of PM10. The city has however appealed against the decision.
A third example comes from the United Kingdom, where several cities are struggling with the NO2 limits. Because 40 (out of 43) air quality zones were exceeding the limits in 2010, the Commission required action plans to be drafted to show how they were going to ensure compliance by 2015 at the latest. After reading the draft action plans, Client Earth, an environmental law organisation, noted that 17 of them did not demonstrate sufficient measures and did not expect to be in compliance until 2020, and in the case of London not until 2025.
In late July 2011, Client Earth decided to launch a judicial review against the UK Secretary of State for Environment, Food and Rural Affairs for failing to protect the health and lives of people through the inadequate NO2 plans. However, a High Court judge dismissed the case at a hearing in London in December, saying that it was up to the European Commission, not a national court, to take corrective action.
The national conditions for pursuing legal cases to improve air quality vary considerably across member states. There are several other countries where, as in Germany, environmental organisations cannot be stakeholders. A recent precedent-setting ruling in the ECJ might however have changed these circumstances completely.
A Slovakian environmental group got the right to be party in a case concerning habitat protection for bears, through the Aarhus convention, which the European Union has signed and ratified. This can probably, though it has not been tried yet, open up the right for environmental groups to be a party in court cases concerning national law that follows under the EU air quality legislation.
The financial risk for the pursuer is another factor that varies. In some countries there is a risk of having to pay all trial costs (which can amount to hundreds of thousands of euro), while in other countries trial costs are either paid by the state or there are other legal devices that protect financially weaker parties.
One way to avoid these costs is to threaten to pursue legal action, put the air pollution problem in the public eye and hope that the resulting pressure will be enough to settle an agreement with the local authority. This strategy was proven successful in Denmark in 2008, when four environmental groups prepared a lawsuit against the Danish government for not doing enough about PM10 levels in the capital. This caused headlines in some of the leading newspapers, and the Environmental Protection Agency subsequently came up with the necessary action plans.


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U Minnesota Morris Achieves Electricity Carbon Neutrality
Roughly 70 percent of the campus' electricity is generated by renewable energy–two university-owned, 1.65-megawatt wind turbines and several photovoltaic systems. To achieve carbon neutrality, the campus recently purchased renewable energy credits equal to the fossil-fuel produced electricity the campus uses.
U Pittsburgh Signs Solar PPA for Electricity
Through a new, 20-year solar power purchase agreement, the university will purchase all of the renewable electricity produced by a planned 20-megawatt solar power facility. It will cover approximately 13 percent of the Pittsburgh campus' annual electricity usage.
Purchasing News
AASHE Purchasing Resources
Queen Mary U Invests in Energy Efficiency Measures
(U.K.) The university will be installing energy monitoring, enabling remote accessibility and live energy reporting, and LED lighting systems. The changes are expected to cut energy use by 25 percent at its Whitechapel campus.
South Dakota State U Adds 20 KW Photovoltaic Array
In early June, 54 solar panels were installed on the university's Facilities & Services roof. The 20.25-kilowatt solar-electric array provides approximately 18 percent of the total building energy usage.
Paul Smith's College Receives $60K for Energy Audit & STARS Fellows
The New York State Energy Research and Development Authority (NYSERDA) recently awarded a $56,000 REV Campus Challenge Technical Assistance for Roadmaps grant to the university that will be used to hire third-party energy consultants to assess campus infrastructure and energy use. Additionally, $4,000 in bonus funding will support sustainability fellowship positions for students working on Sustainability Tracking and Rating Systems (STARS) projects.
U Virginia Partnership Increases Adoption of Solar Energy
In an effort to help the state reach its carbon neutrality goal of 2050, the University of Virginia Weldon Cooper Center for Public Service has partnered with the state's Department of Mines, Minerals, and Energy to host an advisory program, SolSmart, aimed to help local governments implement and embrace solar energy.
Bethel College Installs Solar Carport
The new photovoltaic carport adds 52,000 watts to the college's existing solar capacity and includes two electric car charging stations.
DePauw U Installs Photovoltaic Array
The roof of the university's Indoor Tennis and Track Center now holds 860 photovoltaic panels. More than 40 donors gave toward the installation of the panels, which cost $460,000.
U Toledo Installs 332 KW Solar Array
A new 332-kilowatt photovoltaic array on the university's Health Science Campus is expected to save the university approximately $30,000 per year in avoided electricity costs. A senior student design team worked with the university's Facilities and Construction to identify the site and prepare construction engineering drawings. The Student Green Fund approved spending $350,000 to cover the costs to install the array.
South Dakota State U Installs 7.4 KW Photovoltaic System
A new 7.4-kilowatt system was recently mounted on the university's American Indian Student Center.
North Carolina State U Installs Thermal Energy Storage Tank
Equivalent to a seven-story swimming pool holding more than 3 million gallons of water, the thermal energy storage tank enables the university to create chilled water for campus buildings at night when electricity rates are lowest and store the chilled water for use during the day when electricity rates are highest. The system is designed to supply chilled water for mechanical systems at more than 20 buildings.
Greenville U Completes Solar Installation
The university's recently completed photovoltaic project will cover approximately 75 percent of the university's electricity needs. The project was feasible thanks to an Illinois renewable energy program that allowed the school to acquire the system at no up-front cost and pay over six years with the money that would have been spent on electricity.
Northwestern U to Support 16 Community Solar Projects
The university recently signed an agreement with a community solar provider to support the development and construction of 16 community solar projects in Illinois, totalling 11.9 megawatts. Additionally, the partnership will provide hands-on experiential learning for students. Community solar allows customers to subscribe to a local solar project, receive credits on their utility bill and reduce their overall electricity costs.
George Washington U Installs Solar on Five Buildings
Five buildings at the university's Foggy Bottom campus now house solar panels with a combined capacity of 497 kilowatts.
Macquarie U Signs Hydroelectricity Agreement
(Australia) The new seven-year contract allows the university to source hydroelectricity for its North Ryde campus. Under the contract, the university will purchase approximately 54,422 megawatt-hours of electricity per year.
Emory U to Install 5.5 MW of Solar
The university will install more than 15,000 solar panels across 16 buildings on its Druid Hills campus, which will generate approximately 10 percent of Emory's peak energy requirements. The project is part of a 20-year agreement to install 5.5 megawatts (MW) of solar electricity across campus.
John Wood CC to Install Photovoltaics
The college expects the installation of a new photovoltaic system on the Agricultural Education Center to be completed in mid-June 2020. The project was partially funded by a grant from the Illinois Green Economy Network, an AASHE Affiliate.
Energy Dept. Announces Solar Decathlon Winners
For the first time, the U.S. Department of Energy Solar Decathlon Design Challenge recognized and awarded two grand winners. The winners were University of Oregon in the commercial building division and Miami University in the residential division. The U.S. Department of Energy Solar Decathlon is a collegiate competition that challenges student teams to design and build highly efficient and innovative buildings powered by renewable energy.
U New Mexico to Install Solar Array on Library
A joint effort between the Associated Students of UNM and UNM's Facilities Management has resulted in approval of a $400,000 solar array to be built atop the university's Zimmerman Library. With an estimated peak output of 429 kilowatts, the solar array is expected to meet the electricity needs of the library. Any surplus electricity will be fed onto the grid. The university is expected to save $25,000 per year in avoided costs for electricity.
Villanova U Procures Hydroelectric Power
In a new multi-year renewable electric energy agreement, the university will receive approximately 26,000 megawatt-hours of hydroelectricity.
Cornell U Connects 18 MW Solar Farm
In mid-January, the university flipped the switch on its largest solar project, totaling 18-megawatts. The new solar farm will generate approximately 20 percent of Cornell's electricity use.
Arizona State U Achieves Carbon Neutrality
The university has recently achieved net zero greenhouse gas emissions from campus operations, six years earlier than its commitment of 2025. Increased energy efficiency in both new buildings and campus retrofits; on-site solar generation; renewable energy purchases from large-scale, off-site generation facilities; and purchase of carbon offsets and renewable energy were deployed. The carbon neutrality commitment date for emissions related to commuting and air travel is 2035.
Allegheny College Becomes Carbon Neutral
Allegheny set a goal of achieving carbon neutrality by 2020 as part of a climate action plan adopted by the college's board of trustees in 2010. Three key efforts contributed to the college's achievement of this goal: reducing emissions through implementing efficiency measures; decarbonizing energy sources by shifting from fossil fuel to renewable energy; and offsetting emissions through projects that offset or remove carbon.
U Pennsylvania Signs 220 MW Solar PPA
Consisting of two contracts, the agreement will result in two new solar energy facilities with combined capacity of 220 megawatts, which will produce an estimated 450,000 megawatt-hours of electricity annually. The purchased electricity will be equal to approximately 75 percent of the total electricity demand of the academic campus and Penn Medicine, the University of Pennsylvania Health System.
Dickinson College Achieves Carbon Neutrality
Dickinson recently achieved its carbon neutrality goal by cutting its greenhouse gas emissions 25 percent from 2008 levels and purchasing carbon offsets to mitigate the remaining emissions. From 2020 forward, all the activities included in the college's carbon footprint—like heating and cooling the campus, use of vehicles and air travel—will result in zero net emissions of greenhouse gases.
Northwestern U Receives Sustained Excellence Award
The university recently received the U.S. Environmental Protection Agency's 2020 Energy Star Partner of the Year - Sustained Excellence Award for energy efficiency and energy conservation solutions.
Cornell U Releases Lab Shutdown Procedures
The university recently released a set of guidelines to assist lab managers in quickly identifying energy-saving measures to be implemented during the COVID-19 crisis.
Stony Brook U & U Massachusetts Amherst Win Energy Award
The two universities were recently highlighted as Customer Project Award winners of the 2020 Smart Energy Decisions Innovation Awards for exemplary use and implementation of innovative technologies and progressive practices that reflect new approaches to energy procurement and management. Stony Brook won for its data analytics platform and UMass Amherst was recognized for its 1.3-megawatt battery storage.
Emory U Receives Energy Dept Smart Labs Award
In the fall, the university received the Accelerating Smart Labs Award for Program Development and Implementation from the U.S. Department of Energy for having developed a comprehensive strategy based on the goals of the Smart Labs program. Some strategies include identifying lab energy conservation measures, estimating savings, seeking funding to expand the program, and starting to implement projects in seven of its lab buildings.
U Southern California Installs Solar on Athletics Arena
The university's Galen Center will soon house 1,500 solar-electric panels that are expected to supply as much as 15 percent of the center's electricity.
Utah State U Approves Recommendations to Lower GHG Emissions
The university recently committed to the following strategies to reduce greenhouse gas emissions: dedicate $60,000 annually in ongoing funds to allow USU Facilities to purchase a renewable energy portfolio from an external power provider; facilities will invest $500,000 each year toward improving building energy efficiencies, and additional funding has been allocated within existing budgets for LED light conversions over the next two years; create a $10 per trip carbon fee on all university-sponsored airline flights, from which funds raised by this fee will be used to invest in on-campus projects that reduce USU's carbon footprint; and enhance academic programs to raise awareness of sustainability and climate change.
Cornell U Signs PPA for 1 MW Rooftop Solar
In December, Cornell finalized a 1-megawatt power-purchase agreement to construct rooftop solar arrays on all five buildings of the North Campus Residential Expansion project. The solar power from this project will reduce the university's carbon footprint by providing about 35 percent of the power needed for the new buildings.
Four Pennsylvania HEIs Partner on Renewable Energy Purchase
Lehigh University and Dickinson, Lafayette, and Muhlenberg Colleges recently signed a 15-year power purchase agreement for the production by a 45.9-megawatt share of a new 200-acre solar farm located in Texas. The partnership will enable all four institutions to mitigate 100 percent of their carbon emissions associated with electricity use.
U Pittsburgh Bradford Residence Hall Earns LEED Silver
The university's newly certified Alexander House features renewable energy, high-efficiency HVAC systems, water-efficient plumbing fixtures and low-VOC furnishings. The building has bike storage for students and parking places for electric vehicles.
U Pittsburgh Renovation Receives LEED Silver
Clapp Hall, the building that houses the department of biological sciences, features energy-efficient lighting, water-saving plumbing fixtures, a demand-control ventilation system and a lab exhaust energy-recovery system. More than 95 percent of construction waste was diverted from landfills, 90 percent of existing building elements were reused, and 36 percent of building materials were regionally manufactured. Clapp Hall also has bike storage and changing rooms for commuters.
Vassar College to Purchase 100% Renewable Electricity
In a new contract with its energy supplier, the college plans to purchase electricity generated solely by renewable sources, effective July 2021.
Michigan State U to Install 20 MW Solar Array
The university's board of trustees recently approved a 20-megawatt solar-electric array that will span nearly 145 acres. The project is expected to cost about $2.3 million and will be funded with university utility reserve funds. It is estimated that the array will begin producing power by the end of 2022 and save the university at least $27 million in the next 25 years.
EPA Releases Top 30 List of Green Power Users
The U.S. Environmental Protection Agency's Top 30 College & University list highlights the largest green power users among higher education institutions within the Green Power Partnership. The combined green power use of these organizations amounts to more than 3.5 billion kilowatt-hours of green power annually.
Colorado State U Pueblo Signs Solar Power PPA
The university recently signed a power purchase agreement (PPA) for a $17 million solar project that will be the main electricity source for the campus.
Clarkson U Purchases Electricity From Hydro
In its effort to reach net-zero greenhouse gas emissions by 2025, the university signed a new agreement to purchase over half of its electricity needs from hydro energy.
William & Mary Signs Power Purchase Agreement
The recently signed 20-year power purchase agreement is for the output of a 20-megawatt photovoltaic system that is projected to generate approximately 50 percent of the university's electricity needs. The production of solar energy at the farm will likely begin in 2021. William & Mary expects the contract will begin to save the university money after about six years.
Colorado College Achieves Carbon Neutrality
The college recently achieved carbon neutrality through a variety of initiatives that include efficiency upgrades, building renovations, campus engagement initiatives, on-site renewable energy, local renewable energy purchases, and carbon offsets.
U Virginia Sports Field & Stadium Earns LEED Silver
The university's Davenport Field and Stadium recently completed a renovation that resulted in the LEED Silver certification from major improvements to the energy systems.
Columbia U Staff Complete Energy Efficiency Training
Forty-five Columbia residential operations and maintenance staff completed the 30-hour New York City Building Operator Training program to help improve energy efficiency of residential buildings and reduce operating costs. The curriculum provides participants with energy and resource conservation techniques related to a building's heating, electrical and water systems.
Marymount Manhattan College Purchases 100% Renewable Energy
The college now purchases all of its power from renewable sources. Additionally, after consulting with an energy and water management company, the college has installed a building management system for heating, cooling and electricity, and installed energy-efficient windows in its Carson Hall.
Ohio U Building Scores LEED Gold Certification
Ellis Hall, a 115-year-old building, was recently renovated and now features LED lighting, low-flow toilets and sinks, new insulation, new windows, and an energy-efficient heating and cooling system. Exterior landscaping includes drought-tolerant trees and shrubs.
U Iowa Enters Public-Private Energy Partnership
The University of Iowa recently received approval from the board of regents to enter into a 50-year, nearly $1 billion partnership with Engie North America and Meridiam. Under the agreement, the university will maintain ownership of the UI utility system and Engie will perform all operations with respect to steam, cooling, water and electricity. Engie and Meridiam will work with the university to ensure that the university meets its goal of being coal-free by Jan. 1, 2025, if not sooner; that sources of renewable fuels and incorporating sustainable, lower-cost fuel options into the existing utility systems will be explored; and that experiential learning and research opportunities are available to students, faculty and staff.
Colgate U Buildings Earn LEED Gold
Two new residence halls, Jane Pinchin Hall and Burke Hall, recently acquired LEED Gold status.
Selkirk College Installs Solar Array
In October, the college installed a solar array on the roof of the library that includes 275 panels with the capacity to produce an estimated 112,000 kilowatt-hours of electricity per year. The project will produce approximately 10 percent of the electricity needed on an annual basis at the college's Castlegar campus.
Pace U Installs Solar Tree
The new tree-shaped design holds seven solar panels and is nearly 15 feet tall, providing seating for six, wireless internet, six USB ports and an interactive display.


text:
package me.hulipvp.chambers.command.commands;

import me.hulipvp.chambers.koth.structure.Koth;
import me.hulipvp.chambers.command.ChambersCommand;
import me.hulipvp.chambers.util.commandapi.Command;
import me.hulipvp.chambers.util.commandapi.CommandArgs;
import net.md_5.bungee.api.ChatColor;

public class KothCommand extends ChambersCommand {

	@Command(name = "koth", description = "See info about the Koth", aliases = { "checkkoth" }, usage = "/<command> <args>", playerOnly = true)
	public void onCommand(CommandArgs commandArgs) {
		Koth koth = plugin.getKothManager().getKoth();
		if (koth == null) {
			commandArgs.getPlayer().sendMessage(plugin.getKothManager().getKothFormat() + ChatColor.UNDERLINE + plugin.getGameManager().getGame().getKothName() + ChatColor.RESET.toString() + ChatColor.YELLOW + " is not running.");
		} else {
			commandArgs.getPlayer().sendMessage(plugin.getKothManager().getKothFormat() + ChatColor.UNDERLINE + plugin.getGameManager().getGame().getKothName() + ChatColor.RESET.toString() + ChatColor.YELLOW + " is able to be contested.");
		}
	}

}

redpajama_set_name: RedPajamaGithub


Hii. 🙂 Thanks for your support and encouragement, expressed by your visits, likes, comments and follows.
However, in view of the fact that I am still getting blog and email followers after my last post, where I announced my movement to a self-hosted WordPress blog @ www.infoplaze.com, I decided to do this post.
In this regards, I want to really thank you for the follow and visit[s] and would like to inform you [again] that I now blog at www.infoplaze.com. That means that you won't get new posts here. So, I am passionately begging you for your continued support by way of your very encouraging followership, visits, likes and comments at my new blog.
Once more, thanks a lot for your encouragement.
I look forward to having you around.
Infoplaze.com, content-wise, promises to be better, more informative, educating and inspiring. Since it is a self-hosted site, I am working hard to make sure it lives up to its name…INFOPLAZE.
So I humbly urge you to follow me or subscribe to my posts by email @ http://www.infoplaze.com and let us continue from where we left off.


text:
Q: Chart js not display graph I use chart js to display my graph, but it wont displaying it...
I try to check it in jsfiddle and it works there, but on my website it actually wont work.
The source from chart.js is correctly embedded, so i don't know what the error could be
jsfiddle Link
My code:

var diemeltalsperrectx = document.getElementById('diemeltalsperre').getContext('2d');
var diemeltalsperre = new Chart(diemeltalsperrectx, {
  type: 'line',
  data: {
    labels: [
      '08.05.2020 - 18:15:00',
      '09.05.2020 - 01:00:00',
      '09.05.2020 - 06:15:00',
      '09.05.2020 - 18:15:00',
      '10.05.2020 - 01:00:00',
      '10.05.2020 - 06:15:00',
    ],
    datasets: [{
      fill: true,
      label: 'Pegelstand (cm)',
      data: [
        374.91,
        374.9,
        374.9,
        374.87,
        374.87,
        374.84,
      ],
      backgroundColor: [
        'rgba(0,0,128, 1)'
      ],
      borderColor: [
        'rgba(0,0,128, 1)'
      ],
      borderWidth: 3
    }]
  },
  options: {
    scales: {
      yAxes: [{
        ticks: {
          beginAtZero: true
        }
      }]
    }
  }
});
<script src="https://cdnjs.cloudflare.com/ajax/libs/Chart.js/2.3.0/Chart.js"></script>
<canvas id='diemeltalsperre' width='800' height='400'></canvas>

A: The values are very close to each other, try with:
beginAtZero:false

redpajama_set_name: RedPajamaStackExchange


text:
Former Liverpool coach 'thrilled' after his team do a job on Sam Allardyce
Neil Critchley's Blackpool 'play the Liverpool way' to do a job on Sam Allardyce's West Bromwich Albion in the FA Cup
Chris Beesley
Get the biggest Liverpool stories with our LFC newsletter
Former Liverpool coach Neil Critchley did a job on Sam Allardyce as League One Blackpool rocked Premier League West Bromwich Albion in the FA Cup.
The sides drew 2-2 after extra time at Bloomfield Road in their third round tie on Saturday but the Tangerines recorded a famous giant-killing scalp by triumphing 3-2 in the resultant penalty shoot-out.
Given the way Allardyce crowed following his side's 1-1 draw with Liverpool last month – maintaining his unbeaten record against Jurgen Klopp at Anfield – playing an ultra-defensive 6-4-0 formation, declaring that "I've been able to master that over many years", Kopites may be encouraged to learn the manner in which Blackpool achieved their victory.
Liverpool expected to lose £42m after Man United and Tottenham financial blow
Manchester United set for triple injury boost ahead of Liverpool showdown
Managed by Critchley, who left his post as the Reds Under-23s boss in March last year to be handed his first full-time senior role after previously coaching Liverpool's Under-18s between 2013-17, the Daily Mail reports that he has gradually improved them by imposing 'the Liverpool way' on the team.
Their article states: "Sam Allardyce's West Bromwich could not cope with the crisp, first-time passing of the underdogs, whose goals both came from slick moves that started in their own half."
Since his point against the reigning Premier League champions, Allardyce has endured a miserable run with this cup upset following back-to-back home thrashings against Leeds United (5-0) and Arsenal (4-0).
*Transfer news, rumours and analysis...sign up for The Gossip newsletter.
For Critchley, once one of Klopp's trusted lieutenants though, it was an occasion to savour.
The 42-year-old said: "It's a great day for us, for the club and the supporters.
"We went toe-to-toe with Premier League opposition and more than matched them for entirety of the game.
(Image: LINDSEY PARNABY/AFP via Getty Images)
"To a man, from (goalkeeper) Chris Maxwell with his penalty saves to all of the lads who came on as substitutes, they couldn't have given any more.
"I'm absolutely thrilled to bits. It's a real shame the supporters weren't there with us, but hopefully they're proud of what they saw the team produce.
"We're thrilled to get through to the next round and we'll look forward to the draw on Monday."
Neil Critchley


text:
Q: How to delay data shown on scene? Description:
I have a StackPane as parent container (display) and my components are extending Region and kept in a ConcurrentLinkedQueue. The display of the components are not a problem, even the animation work as I wanted in sync with a timeline (the timeline records the whole process. E.g. min 2 -> Circle pop's up and does some fading + transition, etc.). All nice until now.
Now my problem comes: first of all I want after I consume and play this component to delete it from the display and from the Queue.
Second thing, now when I run this and I have two components that they are at the same time they run together in parallel, even though they are polled from queue (that I don't understand). So I need something that delays or waits until the first component was played, after this delete this component and play the next.
Any suggestion will be awesome!
        //show message
    public void showMessages() {
            while(!messageQueue.isEmpty()) {
                    playMessage(messageQueue.poll());
            }
    }

    /**
     * Searches for the source and target component, finds the connection points and prepares the message to be played
     * @param mess Visual message node, that will appear on dashboard
     * If no source and target component will be found this will return without doing anything
     */
    public void playMessage(MessageNodeVGT mess) {
            //get the source and the target of the message
            CompNodeVGT source=getComponent(mess, SOURCE);
            CompNodeVGT target=getComponent(mess, TARGET);
            if(source==null || target == null) {
                    return;
            }
            Point2D sourcePoint=get2DPoint(source.impl_getShape(),target.impl_getShape());
            Point2D targetPoint=get2DPoint(target.impl_getShape(),source.impl_getShape());
            mess.setSourceCoordinates(sourcePoint);
            mess.setTargetCoordinates(targetPoint);
            instance.getChildren().add(mess.createPath());
            instance.getChildren().add(mess);
            mess.play();
    }

A: Due to the fact that I have two timelines (one general for the whole application and one local for the animation/s) the lock or other technique didn't help. Here it's why:
1st local animation ~ 3seconds (duration)
2nd local animation ~ 3seconds (duration)
general timeline: |-----10s----20s---x-x2---..
let's say x=22s =2200 millis, this is when the 1st animation pops up if the second animation will be at x2 and if this x2 is in between 1 millis - 1000 millis (even more) I will see this as a parallel event, even this is not completely true.
What I did? I just paused the general timeline until the 1st animation finishes, then on setOnFinish I just resume the general timeline.
This is not a perfect answer, because actually with this Pause you lose some time in the case if you are trying to follow some event in Real time.
I'm open to other solutions, hope this helps others too.


text:
Q: Why adding some packages breaks my code? I noticed that adding packages sometimes breaks my code. Functions like Seq.replicate and List.take that were working get red squiggly lines and the program no longer runs. Here is a detailed procedure that should replicate the problem (I am using VS Community Edition 2017):
1) Create a project.
2) Program.fs contains the code:
[<EntryPoint>]
let main argv =
    printfn "%A" argv
    0 // return an integer exit code

3) Add two lines of code as follows:
[<EntryPoint>]
let main argv =
    let repla = Seq.replicate 10 "A"
    printfn "%A" (repla |> List.ofSeq |> List.take 5)
    printfn "%A" argv
    0 // return an integer exit code

3) Hit Ctrl+F5, the program runs without any problems.
4) Right-click on References on the Solution Explorer. Click on Manage Nuget Packages....
5) Click on Browse.
6) Search for Newtonsoft.Json and install it.
7) Hit Ctrl+F5, the program runs without any problems. The console prints
["A"; "A"; "A"; "A"; "A"]
[||]

8) Proceed as in steps 4 to 6 and install package MathNet.Numerics using Nuget.
9) Hit Ctrl+F5, the program runs without any problems. The console prints
["A"; "A"; "A"; "A"; "A"]
[||]
10) Proceed as in steps 4 to 6 and install package MathNet.Numerics.FSharp using Nuget.
11) Click on the Program.fs tab. The dreaded red squiggly lines appeared under Seq.replicateandList.take`.
12) Hit Ctrl+F5. A dialog box pops up with the following message:
There were build errors. Would you like to continue and run the last successful build?

Questions:
a) What exactly is happening?
b) Is this an isolated case related to a problem in MathNet.Numerics.FSharp or the specific combination of packages installed, or the order in which they were installed? Or is this a common problem?
c) Is it possible to avoid this problem while using Nuget?
d) If not, is it possible to avoid this problem by installing by some other means (not Nuget)?

A: MathNet.Numerics.FSharp has a dependency to FSharp.Core.3.1.2.5 which replaces the current version of FSharp.Core you are using. Being quite old 3.1.2.5 lacks many functions.
This happens with other popular F# libraries such as FsCheck as well. I usually change the reference to the newest version of FSharp.Coreas that should be backward compatible.
To do so I unload the F# project and update the FSharp.Core reference in the project file to:
<Reference Include="FSharp.Core">
  <Name>FSharp.Core</Name>
  <AssemblyName>FSharp.Core.dll</AssemblyName>
  <HintPath>$(MSBuildProgramFiles32)\Reference Assemblies\Microsoft\FSharp\.NETFramework\v4.0\$(TargetFSharpCoreVersion)\FSharp.Core.dll</HintPath>
</Reference>

Then I reload the project.
As this is a rather clunky procedure hopefully more insightful users will post a better solution.

A: Well, I think the OP's concerns are somewhat valid. I would discourage hand-editing the fsproj file. And also the comments are very valid too, Paket is a great tool that can simplify dependency management both in VS and Code. So here's a very simple, two part answer, where a) you can actually get your solution working using nugget and without hand-editing the project file, and b) get a quick intro to how using paket on VS.
This issue where some package downloads and old dependency that messes up other code occurs now and then and is probably due to a strategy of relying on the minimum viable version. Your issue is somewhat similar to this Q: Why can't I get suave to work.
Here's the simplest solution using just nugget:

*

*Open a new solution with an fsharp console project

*At this point your Fsharp.Core in VS2017 will be 4.1:

*Now add Mathnet.Numerics.Fsharp via nugget:

Now, very unfortunately you got downgraded to F# 3.1

*This is no good, so just get Fsharp.Core via nugget as well!

*And voila, you're back to a working solution without needing to edit fsproj. Basically all you had to do is add the Fsharp.Core package. You could have also changed the .NET Framework option in properties and move it to .NET 4.7 with Fsharp 4.1:

Now for Part 2 of the answer with Paket (btw, the Paket plugin just got updated and I tested and it works fine). You can essentially use it as a replacement for nugget or directly edit the files. You actually need to work with two files, paket.dependencies in the solution root and paket.references in the project root. Paket.lock will be generated. Here's a two-step process to get started. Take the project you just made above and from the Tools menu in VS select Paket Dependencies Manager, do Initialize Paket, then do Convert From Nuget:

With this you can keep managing your dependencies with nuget or use paket if you like. Next I will add Newtonsoft.JSON to the references. Double-click on the paket.dependencies file and add the following line: nuget Newtonsoft.Json 10.0.3 restriction: >= net452 , you only actually need Newtonsoft.Json in there but we don't want to download the whole interweb called .NetCore. Also add this line to paket.references (you can just click on it in the project explorer): Newtonsoft.Json. And the run Tools | Paket | Install:
Paket.dependencies:

Paket.references:

And Run Paket Install:

And you will have Newtonsoft.JSON installed. Usually you don't need to specify a version of framework restriction, however it has some pre-release package that you might not want, and also I assume you don't need the .netcore dependencies.
You can actually right click and try install this package from the references Add Package menu, but you will bump into dependency errors.