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Обычная тренировка в спортзале для сравнения:
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Сначала 10 мин. разминки на элептическом, потом обычные тренажеры, круто видно сколько подходов было сделано =)
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Quote:
охренеть!!! даешь фото топлес!!!
развивай воображение
В ответ на:
среди семестра был забавный случай с одним студентом по фамилии Погодаев, который не дошел ко мне на лекцию (это была первая пара) и лег спать на диванчик в коридоре. Его растолкал охранник спросил кто такой и откуда и привел его ко мне на лекцию. Студент пожаловался, что очень хотел спать, но злобная родительница выгнала его на занятия
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We shall summarize the whole procedure. For
details one should refer to the original papers [35,
41]. Both ventricles from one pig heart were cut into
small pieces with scissors after removal of the fat and
of the main coronary arteries. The tissue was then
resuspended in 3.5 vol of buffer I (10 mM histidine,
free base, pH 7.5, 750 mM NaC1, 1 mM PMSF),
mildly homogenized (low speed, 5 s) and centrifuged
at 14 000 x g for 20 min. The supernatant (SI) was
discarded. This procedure was repeated twice, first
with the same buffer and then using buffer II (5 mM
histidine, 10 mM NaHCO3, pH 8.0, 1 mM PMSF).
The aim of these 3 mild homogenizations is to remove
most of the sarcoplasmic reticulum with the discarded
supernatants $1, $2 and $3. This goal, indeed, was
achieved, since the specific activity of the Ca 2+ / K +-
ATPase was about 3 times lower in the third pellet
(P3) than in the homogenate (Table I). P3 was then
resuspended in 3.5 vol of buffer II and vigorously
homogenized (high speed, 3 x 30 s) in order to cause
the formation of sealed sarcolemmal vesicles. Nuclei
and cell fragments were then sedimented by centrifugation
(14 000 x g, 20 min) and discarded. At that
stage, the specific activity of the Na + / K+-ATPase in
the supernatant ($4) was 5 times higher than that in
the whole homogenate. The microsomes present in
$4 were pelleted by centrifugation at 44 000 x g for
30 min. This pellet (P 5) was resuspended in buffer
III (10 mM histidine, free base, pH 7.5, 0,25 M
sucrose) and centrifuged twice, one at low speed (17
000 x g, 20 min), next at highspeed (95 000 x g, 30
min), in order to separate the sarcolemmal microsomes
from the contaminating mitochondria. After the
latter centrifugation, a 2 rnl layer of sarcolemmalenriched
microsomes (CM7) was recovered on top of
a mitochondria-enriched brown pellet, as demonstrated
by the differential partitioning of the
Na* / K+-ATPase and succinate dehydrogenase activities.
The sarcolemmal vesicles were further purified
by centrifugation of CM7 on a 0.6 M sucrose cushion
(90 min, 170 000 x g). The white material collected
at the interface (IF) was washed in 10 vol of
deionized water and centrifuged again at 170 000 ×
g for 90 min. The resulting pellet (SM), consisting of
purified sarcolemmal membranes, was resuspended
in buffer I11. According to the Na*/K+-ATPase
marker, the purification was of the order of 50 times
(Table I).
Очень зонятно и поучительно
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