Internazionale Main Thread II

[Dejan-Stankovic]

[Totally FRANK]

[Messinterista]

[Dejan-Stankovic]

Frank; No way on earth we should give up Khrin, regardless of Vieira leaving or not. If they want youth players, it should be from the Primavera and not players like Stefanovic or Obi.

MI; yeah he's about to hit his peak as a footballer, 26-28, as most people say.

Onto the Genoa game and they've got quite a few players out but so do we. Milito and Motta won't be playing but I'm not too sure about Lucio as of yet. Maicon is also back after the suspension so we should still field a decent team.

-------------------Cesar--------------------
Maicon-----Lucio--------Samuel----Chivu
----------------Cambiasso-----------------
------Zanetii---------------Stankovic------
----------------Sneijder--------------------
----------Eto'o------------Balotelli---------

This is going to be a tough one, especially since we have our most important game of the season vs Dynamo immediately after. I watched Dynamo vs Barca at the Nou Camp and for the first 20 mins, they were all over Barca so I'm expecting nothing short of a tough game.

[Lombardiamia]

I might be wrong, but last I've heard about Lotito's expectations were something around 4million + 2 player ON LOAN. In fact, there was speculation that Suazo was one of those players wanted by Lotito, whereas we are offering 2 youngsters. No way would be giving Suazo + another player + cash for a player who will be a free agent in a couple of months. Therefore, there could be a possibility of Khrin leaving in january, and I cant say it would be a bad idea either, considering it will be a loan move.
With regards to the Genoa match, they are though against anyone in their stadium. They might have some absentees, but they have such an organized team movement that sometimes it makes it irrelevant whether 1 or 2 players are missing. We will suffer and it will be tough, extremely tough. Let's hope we can capitalize our chances for once and not have to waste 7-8 chances to net 1. Also, I'm quite eager to see Balotelli playing as a 'seconda punta' and Eto'o hopefully playing as a 'prima punta', instead of seeing these guys wasted on the flanks.

[Dejan-Stankovic]

Agreed. Eto'o should always be as close to goal as possible. With Milito-Eto'o, there has to be some sacrifices since both of them are primarily finishers but when Balotelli plays, he should be the one picking up the ball and running, much like Zlatan, although still not as technically sound. It would be a bloody good time for Balotelli to score his first goal of the season in either of the next two games...

[zafferano]

Same doubt on Eto playng , looks like he have a foot injury.
and about pandev 4 mil plaus 2 players , is all media bulshit .
chance he get out of is contract and we will have him for free.
dejan are you fat ?
suffer from colesterol ?
the cura on the way to aussyland in january o febraury .
lol

[Lombardiamia]

[Totally FRANK]

Eto'o foot injury hurts Inter
Thursday 15 October, 2009
Inter have something of an injury crisis up front after Samuel Eto'o suffered a badly bruised foot while on international duty.

Initially it was feared the Cameroon international had fractured something, but a scan showed that nothing was broken.

The 28-year-old is a doubt for this weekend's match against Genoa and could even miss Inter's Champions League tie with Dynamo Kyiv.

Jose Mourinho is particularly worried about that eventuality as Diego Milito is still recovering from a muscle complaint and Mario Balotelli is suspended after being sent off away to Rubin Kazan.

A friendly against Piacenza at the weekend also revealed that Marko Arnautovic is struggling for fitness, as he couldn't play the full 90 minutes.

Inter are looking to reinforce in January with Lazio striker Goran Pandev at the top of their wish list.

However, in the meantime Mourinho might have to either rush players back or come up with a makeshift strike partnership.

[zafferano]

[Messinterista]

[Dejan-Stankovic]

[Dejan-Stankovic]

[zafferano]

Hypolipidemic Effects of Citrus bergamia Risso et
Poiteau Juice in Rats Fed a Hypercholesterolemic
Diet
NATALIZIA MICELI,*,† MARIA R. MONDELLO,† MARIA T. MONFORTE,†
VASILEIOS SDRAFKAKIS,† PAOLA DUGO,# MARIA L. CRUPI,§ MARIA F. TAVIANO,†
RITA DE PASQUALE,† AND ADA TROVATO†
Pharmaco-Biological Department, University of Messina, Vill. SS. Annunziata, 98168 Messina, Italy;
Pharmaco-Chemical Department, University of Messina, Vill. SS. Annunziata, 98168 Messina, Italy;
Department of Organic and Biological Chemistry, University of Messina, Sperone 31,
98166 Messina, Italy
Citrus bergamia Risso et Poiteau fruits have been traditionally utilized, in Calabria (Italy), as a popular
remedy for their hypolipidemic properties. C. bergamia juice total phenol content (2474.35 ( 38 μg/
mL) was evaluated by the Folin–Ciocalteu method; moreover, HPLC analysis led to the identification
of naringin (520 ppm), neoeriocitrin (370 ppm), and neohesperidin (310 ppm). The present study
was designed to investigate the hypolipidemic effects of C. bergamia juice and its protective effect
on liver of hyperlipidemic rats. Chronic administration of C. bergamia (1 mL/rat/day) provoked a
significant reduction in serum cholesterol, triglycerides, and low-density lipoprotein (LDL) levels and
an increase in high-density lipoprotein (HDL) levels; moreover, histopathological observations showed,
in rats submitted to C. bergamia treatment, a protection of hepatic parenchyma. In addition, fecal
neutral sterols and fecal bile acid excretion was found to be increased after C. bergamia treatment.
These results suggest that the hypocholesterolemic effect of C. bergamia may be mediated by the


Metabolic Syndrome, Cholesterol, Diabetes,
Cardiovascular Health, Weight Los

increase in fecal neutral sterols and total bile acids excretion. In addition to the hypolipidemic effect,
the juice shows radical scavenging activity in the diphenylpicrylhydrazyl (DPPH) test; probably the
two effects are related. These observations suggest that the positive intake of C. bergamia may
reduce the risk of some cardiovascular diseases through its radical scavenging function and
hypocholesterolemic action.
KEYWORDS: Citrus bergamia juice; flavonoids; hypolipidemic activity; fecal bile acids; DPPH test
INTRODUCTION
Citrus bergamia Risso et Poiteau (bergamot) is a small tree
belonging to the family Rutaceae. The fruit is pale yellow,
pyriform, and 7.5–10 cm in diameter (1).
C. bergamia appeared in southern Italy before 1700 and is
defined as a hybrid of Citrus aurantium L. and Citrus limon L.
Burm. Fil., by some authors, or of C. aurantium L. and Citrus
aurantifolia (Christm.) Swing. by others (2).
To date, 95% of worldwide bergamot production occurs in
the Ionic area of Calabria (Italy), where soil characteristics and
pH (<6.5–7.5) are particularly suitable for its cultivation (3).
Particularly, the province of Reggio Calabria yields 55% of the
production, and the remaining part comes from the Calabrian
Ionic coast (4). The annual Italian production of bergamot
amounts to 25000 tons (5).
C. bergamia fruit is used mostly for the extraction of its
essential oil from the peel, which is widely used in the cosmetic,
pharmaceutical, and food industries (5). C. bergamia juice,
obtained from the endocarp after essential oils extraction, is
considered to be just a secondary and discarded product (6).
Recently, some authors have reported information on the
flavonoid composition of C. bergamia juice (7, . It is wellknown
that neoeriocitrin, naringin, and neohesperidin are the
flavanone-O-glycosides to be found in the highest amounts in
bergamot juice (257.0–295.8, 248.1–274.6, and 206.6–235.7 mg/
L, respectively) (8). The same authors have identified another
eight flavonoids, lucenin-2, stellarin-2, isovitexin, scoparin,
orientin 4′-methyl ether, rhoifolin 4′-O-glucoside, chrysoeriol
7-O-neohesperidoside-4′-O-glucoside, and chrysoeriol 7-Oneohesperidoside,
in bergamot juice (8).
Bergamot juice, moreover, is characterized by the high
content of fatty acids constituting 79.50% of total acids [oleic
* Author to whom correspondence should be addressed (telephone
+39-090-6766530; fax +39-090-6766474; e-mail nmiceli@
pharma.unime.it).
† Pharmaco-Biological Department.
# Department of Organic and Biological Chemistry.
§ Pharmaco-Chemical Department.
J. Agric. Food Chem. XXXX, xxx, 000 A
10.1021/jf071772i CCC: $37.00  XXXX American Chemical Society
Published on Web 11/27/2007
acid (33.98%), linoleic acid (24.00%), and palmitic acid
(15.34%)], by an acidity value of 0.710 g/L, and by a total sugar
content of 72.760 g/L (2).
Previous studies have shown that a chronic administration
of some Citrus species fruit juice positively influences plasma
lipid levels and can be associated with reduced risk of coronary
heart disease (9, 10). Literature data show that the hypolipidemic
effects can be correlated to several components of Citrus juices,
such as flavonoids (naringin and hesperidin), pectins, and
ascorbic acid, which have a high antioxidant potential and
interfere with cholesterol metabolism (10–12).
Few studies have focused on the biological activities in C.
bergamia juice; in Calabria the fruit juice has been traditionally
utilized as a popular remedy for its hypolipidemic properties
(6). However, the scientific basis for this use in folk therapy
has not been examined. The present study was designed to
investigate the hypolipidemic effects of C. bergamia juice and
its protective effect on liver of hyperlipidemic rats.
MATERIALS AND METHODS
Plant Material. C. bergamia Risso et Poiteau fruits were collected
from plants growing in a cultivation located in Mortara di Pellaro
(Reggio Calabria, Italy).
Preparation of Juice. Bergamot juice was obtained, as a blend of
three cultivars, ‘Fantastico’ (95%), ‘Femminello’ (5%), and ‘Castagnaro’
(5%), by hand-pressing; after it was filtered, small aliquots
(25 mL) were stored at -20 °C and defrosted immediately before the
experiments.
Determination of Total Phenolic Content. The phenol content was
measured by using the Folin–Ciocalteu (reagent) method (13). One
hundred microliters of C. bergamia juice, previously centrifuged (4
°C, 10 min, 3000 rpm), was mixed with 0.2 mL of Folin–Ciocalteu
reagent, 2 mL of H2O, and 1 mL of 15% Na2CO3, and the absorbance
was measured at 765 nm, after 2 h of incubation at room temperature,
with a model UV-1601 spectrophotometer (Shimadzu, Milan, Italy).
All reagents were purchased from Sigma-Aldrich. Gallic acid was used
as a standard, and the total phenols were expressed as micrograms of
gallic acid equivalents per milliliter of juice. The value of total phenolic
content was obtained as the average of three determinations, and the
result is expressed as mean ( standard deviation (SD).
Determination of Flavonoids. The HPLC analysis of C. bergamia
juice was performed using a Shimadzu HPLC system (Kyoto, Japan)
equipped with an SCL-10Avp controller, two LC-10ADvp pumps, a
DGU-14A degasser, and an SPD-M10vp diode array detector. The
column used was a Luna C18 (250 × 4.6 mm i.d., 5 μm) purchased
from Phenomenex (Torrance, CA). A mixture of water, acetonitrile,
isopropanol, and formic acid (78:12:10:0.1, v/v) was used as a mobile
phase in isocratic mode. The mobile phase flow rate was 0.8 mL min-1.
Sample volume injected was 20 μL of undiluted bergamot juice,
centrifuged (3000 rpm) and filtered through 0.45 μm membrane filters
before analysis. Data were acquired using a photodiode array detector
in the range of 190–370 nm, and the chromatograms have been extracted
at 283 nm. Time constant was 0.64 s and frequency 1.5625 Hz. Data
acquisition was performed by Shimadzu LC solution software.
Pure standards (neoeriocitrin, naringin, neohesperidin) were purchased
from Extrasynthese (Genay, France), and the flavonoids in the
sample were identified by comparison of retention times and spectra
for each peak with the corresponding flavonoid standard.
In addition, HPLC-ESI-MS analysis was carried out for further
confirmation of peak identification. The HPLC conditions were the same
as for HPLC-PDA analysis.
The following ESI-MS parameters were applied: mass spectral range,
m/z 200–800; interval, 0.2 s; scan speed, 4000; nebulizer gas flow, 1.5
L/min; curved desolvation line (CDL) temperature, 300 °C; CDL
voltage, –34 V; probe voltage, +5 kV; Q array (quadrupole array):
scan; detection gain, +1.5 kV. MS data acquisition was performed by
Shimadzu LabSolutions V2.04 software.
Free Radical Scavenging Activity. Radical scavenging activity was
assayed according to the method of Ohinishi et al. (14). A methanol
1,1-diphenyl-2-picrylhydrazyl (DPPH) solution (0.1 mM) was mixed
with different concentrations of C. bergamia juice (100, 50, 10, 5 μL)
previously centrifuged (4 °C, 10 min, 3000 rpm); the optical density
change at 517 nm was measured for 4 min after the initial mixing with a
model UV-1601 spectrophotometer (Shimadzu). The scavenging activity
was measured as the decrease in absorbance of samples versus DPPH
standard solution. Ascorbic acid (Sigma-Aldrich) was used as reference.
The mean values were obtained from triplicate experiments. Radical
scavenging activity was expressed as the inhibition percentage (15).
Hypolipidemic Activity. Animals. Male Wistar rats (Harlan Italy),
weighing 180–200 g, were used for the experiment. The animals were
kept in standardized conditions (temperature, 22 ( 2 °C; humidity, 60
( 4%; natural light), with water ad libitum. Animal care was in
compliance with Italian regulations on protection of animals used for
experimental and other scientific purposes (D.M. 116192), as well as
with the EEC regulations (Off. J. Eur. Communities 1986, L 358).
Treatment. The effects of C. bergamia juice on lipid metabolism
were evaluated by diet-induced hypercholesterolemia experimental
model in rat (16). The hypercholesterolemic diet (cholesterol, 2%;
sodium cholate, 2%; vitamin mixture, 2%; oligoelements, 0.2%; salt
mixture, 5.8%; coconut oil, 20%; cellulose, 4%; sucrose, 44%; casein, 5%;
drakettprotein, 15%) was supplied by Altromin-Rieper (Bolzano-Italy).
The rats were divided into three groups of 10 animals each:
• Group I (normolipidemic controls) was kept on a standard diet (S.
Morini, Mil rat GLP) for 30 days.
• Group II (hyperlipidemic controls) received the hypercholesterolemic
diet for 30 days.
• Group III received the hypercholesterolemic diet for 30 days; from
the 1st to the 30th day, each rat was administered by gavage with C.
bergamia juice (1 mL/rat).
During the experiment, animals were weighed weekly, and 24 h food
consumption was recorded daily. On day 29, rats were individually
housed in metabolic cages and feces were collected for 24 h.
At the end of the study, the animals were fasted overnight; blood
samples were collected from the carotid artery of the rats under light
diethyl ether anesthesia, and serum was separated and stored at -20
°C until analyzed. Livers and aorta artery were removed and kept at
-80 °C until microscopic observations. Fecal samples were collected
and kept at -20 °C until analyzed.
Serum Lipids. Total cholesterol, high-density lipoproteins (HDL),
low-density lipoproteins (LDL), and triglycerides in serum samples were
determined colorimetrically and enzymatically using commercial assay
kits (BioSystems S.A., Barcelona, Spain). The atherogenic index (AI)
was calculated by using the following equation: (total cholesterol –
HDL-cholesterol)/HDL-cholesterol.
Microscopic ObserVation: Histology. Liver and aorta artery pieces
for histological investigation (5 mm diameter) were fixed in neutralized
4% paraformaldehyde in 0.2% phosphate buffer for 6 h at 4 °C. The
samples were dehydrated in graded ethanols and, finally, embedded in
Bioplast (Biooptica, Milano, Italy). The liver sections (5 μm thick),
obtained by a rotative microtome, were stained with hematoxylin-eosin.
All samples were observed and photographed with Leica DMLD
microscopy.
Fecal Neutral Sterols and Bile Acids Determination. Neutral sterols
and bile acids in the fecal samples were extracted according to the
method of Crowell and Macdonald (17). Feces were freeze–dried for
48 h, weighed, and ground into fine powder. To sample aliquots (200
mg) was added 1 mL of KOH solution, and the mixtures were saponified
by autoclaving at 120 °C for 1 h; after the addition of 1 mL of NaCl
solution, neutral sterols were extracted twice with ethyl ether (20 mL).
The organic phases (upper phases) were pooled, evaporated with a rotary
evaporator, and dried under nitrogen. The lower phase was acidified
with 0.2 mL of concentrated HCl and extracted six times with ethyl
ether (20 mL). The upper phases were pooled, evaporated with a rotary
evaporator, and dried under nitrogen. The concentration of fecal neutral
sterols was determined by gas chromatographic (GC) analysis. Before
the GC-analysis, all samples were diluted 1:10 v/v in n-hexane, and
R-cholestanol was added as internal standard to a final concentration
of 500 ppm.
B J. Agric. Food Chem., Vol. xxx, No. xx, XXXX Miceli et al.

Bergamot juice reduces blood cholesterol levels in
diet-induced hypercholesterolemia in Wistar rats
(Rats with high cholesterol level consuming high
cholesterol diet)
Total
cholesterol
HDL
cholesterol
LDL
cholesterol
Tryglycerid
es
Blood
glucose
Bergamot
juice 20
ml/day/Kg
-42+4% +31 +3% -44+5% -25+% -10+3%

Bergamot polyphenolic fraction (BPF; 500 mg/day for 30 days)
reduces total cholesterol (tChol), LDL cholesterol (c-LDL), tryglicerides (TG)
and blood glucose (BG) in patients suffering from metabolic syndrome


Bergamot extracts normalize metabolic targets
in rats and patients
suffering from dyslipidemia and metabolic syndrome
Vincenzo Mollace and §Franco Romeo
Chair of Pharmacology and Toxicology, University “Magna Graeciaâ€

[Messinterista]

I have faith in Mancini. He will produce some magic given the opportunity. He has underperformed recently though..

[Dejan-Stankovic]

Underperformed recently MI? Since he's been here, I haven't rated him higher than a 5 in any game he's played.

[Lombardiamia]

[Dejan-Stankovic]

[maro]

[Lombardiamia]