A To Z 3rd Lipi

[Frida Kosofsky]

Lipiddroplets (LDs) are composed of neutral lipids such as triacylglycerol & cholesteryl ester that are surrounded by phospholipid monolayers and are seen ubiquitously, not only in adipocytes1). LDs were originally thought to serve as a lipid storage unit, until a recent study showing that LDs play an important role in regulating lipid metabolism, autophagy2) and cellular senescence3). Therefore, LDs have gained great attention as an important tool to elucidate the mechanisms of their formation, growth, fusion, and retraction.

A medium that contained oleic acid (200 μmol/l) was added and incubated overnight. Then, the supernatant was removed and the cells were washed with PBS. Each Lipi product series (1 μmol/l) was added and the cells were incubated for 15 minutes.

HepG2 cells were stained with Lipi-Red and Nile Red. Lipi-Red was imaged with Green excitation (G), but not Blue excitation (B). However, Nile Red was imaged in both filter. Lipi-Red is preferable for multi-staining.

After adding Lipi-Deep Red (0.1 μmol/l) to the Arf4-GFP expressed Hela cells and the cells were incubated for 30 minutes, the cells were treated with 4% PFA (PBS) to fix, and washed with PBS three times. Fluorescent imaging was conducted by confocal microscopy.

After adding Lipi series to the 4% PFA (PBS) fixed mouse liver adipose tissue and the tissue were incubated for overnight, and washed with PBS. Fluorescent imaging was observed by fluorescence microscopy.

Changes in lipid droplets were examined after the addition of oleic acid or Triacsin C (acyl-CoA synthetase inhibitor) to the HepG2 cell culture medium. For analysis, the number and total area of lipid droplets per cell were computed from the images acquired with CQ1, a confocal quantitative image cytometer (Yokogawa Electric Corporation).

CQ1 captured images of lipid droplets with a 447/60 nm bandpass filter and cell nuclei with a 525/50 nm bandpass filter. Lipid droplets and cell nuclei were individually identified and computed the number and total area by using the CellPathfinder analysis software.

Based on the detected data of cell nuclei and lipid droplets, the number and total area of lipid droplets per cell computed were shown in the graphs below. Compared to the control value, the number and total area of lipid droplets per cell were increased 7-10 times by the addition of oleic acid, but the addition of Triacsin C inhibited lipid droplet formation and showed a 50-60% decrease.

Experimental conditions

HepG2 cells (1 x 103 cells) were disseminated on a 96-well plate and incubated overnight. After the culture supernatant was removed, the cells treated with DMEM plus FBS only (control), DMEM plus FBS and 200 μmol/L oleic acid (Oleic acid), and DMEM plus FBS and 5 μmol/L Triacsin C (Triacsin C) were incubated overnight. Cells were then washed twice with PBS buffer, fixed with 4% PFA for 5 minutes at room temperature, and washed twice with PBS buffer again. Finally, cells were stained in the dark for 2 hours at room temperature with 0.5 μmol/L Lipi-Blue working solution, and quantitative analysis was performed through CQ1.

5. Other

Depending on the cell type, lipid droplets can be smaller than usual and it may be too difficult to observe.

If that is the case, please observe under a high magnification microscope or prepare for positive control with oleic acid treated cells.

Lipi means 'writing, letters, alphabet', and contextually refers to scripts, the art or manner of writing, or in modified form such as lipī to painting, decorating or anointing a surface to express something.[2][3]

The canonical texts of Jainism list eighteen lipi, with many names of writing scripts that do not appear in the Buddhist list of sixty-four lipi. The Jaina list of writing scripts in ancient India, states Buhler, is likely "far older" than the Buddhist list.[10]

Lipi means 'script, writing, alphabet' both in Sanskrit and Pali.[12] A lipika or lipikara means 'scribe' or 'one who writes',[13] while lipijnana and lekhā means the 'science or art of writing'.[2][14] Related terms such as lekhā (लेख, related to rekhā 'line') and likh (लख) are found in Vedic[15][16] and post-Vedic[17] Sanskrit texts of Hinduism, as well as in regional languages such as the Pali texts of Buddhism.[18][19]

According to section 4.119 of the Unadisutras as now received, lipi is derived from the Sanskrit root lip.[24] The Unadisutras themselves certainly existed before the time of Pāṇini,[25][26] instances of later interpolations have been raised by Max Mller, although Mller does not discuss whether the sutra related to lipi was interpolated.[25] Salomon in 1995 remarked "The external testimony from literary and other sources on the use of writing in pre-Ashokan India is vague and inconclusive. Alleged evidence of pre-Mauryan writing has in the past been found by various scholars in such sources as later Vedic literature, the Pali canon, the early Sanskrit grammatical treatises of Pāṇini's and his successors, and the works of European classical historians. But all of these references are subject in varying degrees to chronological or interpretive problems."[27]

The Edicts of Ashoka (circa 250 BCE) use the word lipī. According to some authors, the word lipi, which is spelled dipi in the two Kharosthi versions of the rock edicts,[note 1] comes from the Old Persian prototype dipi (????), which also means 'inscription', which is used for example by Darius I in his Behistun inscription.[note 2] E. Hultzsch, an epigraphist in the colonial British Empire, in his 1925 study on the Inscriptions of Asoka, considered the lip derivation untenable because of the two Kharosthi rock edict inscriptions from 3rd century BCE which use dipi instead of lipi. Hultzsch, as well as Sharma, state that this suggests a borrowing and diffusion of lipi from an Old Persian prototype dipi.[24][28][29]

Some Indian traditions credit Brahma with inventing lipi, the scripts for writing.[30] Scholars such as Lallanji Gopal claim some ancient lipi such as the Brahmi script as used in the Indian texts, may have originated in Jainism.[31]

According to Harry Falk, scripts and the idea of writing can be traced to the Indus Valley civilization in the 3rd millennium BCE, but the term lipi in 1st millennium BCE Indian literature may be a loan word from the Achaemenid region, as a variant of Sumerian dub, turned to dipi or dipī.[33] Sanskrit lipi, states Falk, likely arose from a combination of foreign influences and indigenous inventions.[33] One evidence in favor of this view is that the form dipi was used in some of the Kharosthi-script edicts of Ashoka (3rd century BCE) in northwest India (in closest contact to Achaemenid culture) in parallel to lipi in other regions. As dipi was used in Old Persian Achaemenid inscriptions, Hultzsch suggested in 1925 that this proposal is "irresistible."[24] In his theory about the origin of the Brahmi script, Falk states that the early mention by Paṇini could mean that he was aware of writing scripts in West Asia around 500 BCE, and the Paṇini's mention of lipikara may possibly refer to non-Indian writers such as Aramaic scribes.[34]

Falk states that the single isolated mention of lipi by Paṇini, could mean that he was only aware of writing scripts from West Asia around 500 BCE.[34] According to Paul Griffiths, there is "no hard evidence of the use of Brahmi or Kharosthi script" in India before the Ashoka stone inscription, but the climate of India is such as that writing on other materials would not have survived for over 2,500 years. So, states Griffith, "the absence of early witnesses certainly doesn't mean there were none", but there is no "clear textual evidence of the use of writing in the Vedic corpus".[35]

Kenneth Norman (a professor and the president of the Pali Text Society) suggests lipi in ancient India evolved over the long period of time like other cultures, that it is unlikely that a lipi was devised as a single complete writing system at one and the same time in the Maurya era. Norman suggests that it is even less likely that Brāhmī was invented during Ashoka's rule, starting from nothing, for the specific purpose of writing his inscriptions and understood all over South Asia.[36] Reviewing the recent archaeological discoveries relating to writing scripts in South Asia particularly Buddhism, Norman writes, "Support for this idea of pre-Ashoka development [of writing scripts] has been given very recently by the discovery of sherds at Anuradhapura in Sri Lanka, inscribed with small numbers of characters which seem to be Brahmi. These sherds have been dated, by both carbon 14 and thermo-luminescence dating, to pre-Ashokan times, perhaps as much as two centuries before Ashoka".[37]

Jack Goody similarly suggests that ancient India likely had a "very old culture of writing" along with its oral tradition of composing and transmitting knowledge, because the Vedic literature is too vast, consistent and complex to have been entirely created, memorized, accurately preserved and spread without a written system.[38][39] Walter Ong and John Hartley concur with Goody and share the same concerns about the theory that there may not have been any writing scripts during the Vedic age, given the quantity and quality of the Vedic literature.[40]

Richard Salomon, in a 1995 review, states that the lack of securely datable specimens of writing from pre-3rd century BCE period, coupled with chronological and interpretive problems of more ancient Indian texts, has made dating lipi and who influenced whom a controversial problem.[43]

While historical evidence of scripts is found in the Indus Valley civilization relics, these remain undeciphered.[44] There has been a lack of similar historical evidence from the 2nd and early 1st millennium BCE, until the time of Ashoka where the 3rd-century BCE pillar edicts evidence the Brahmi script.[45] Late 20th-century archaeological studies combined with carbon dating techniques at Ujjain and other sites suggest that Brahmi script existed on the ancient Indian subcontinent as early as 450 BCE.[46]

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