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Cunningham, Fisiologia veterinaria, cuarta edición

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Normal Cell Metabolism: Cellular respiration describes the series of steps that cells use to break down sugar and other chemicals to get the energy we need to function. Energy is stored in the bonds of glucose and when glucose is broken down, much of that energy is released. Cell utilize energy in the form of ATP. The first step of respiration is called glycolysis. In a series of steps, glycolysis breaks glucose into two smaller molecules - a chemical called pyruvate. A small amount of ATP is formed during this process. Most healthy cells continue the breakdown in a second process, called the Kreb's cycle. The Kreb's cycle allows cells to “burn” the pyruvates made in glycolysis to get more ATP. The last step in the breakdown of glucose is called oxidative phosphorylation (Ox-Phos). It takes place in specialized cell structures called mitochondria. This process produces a large amount of ATP. Importantly, cells need oxygen to complete oxidative phosphorylation. If a cell completes only glycolysis, only 2 molecules of ATP are made per glucose. However, if the cell completes the entire respiration process (glycolysis - Kreb's - oxidative phosphorylation), about 36 molecules of ATP are created, giving it much more energy to use. IN CANCER CELL: Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful. Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation. This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive. Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful. Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation. This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive. introduction to WARBERG PHENOMENA: WARBURG EFFECT Usually, cancer cells are highly glycolytic (glucose addiction) and take up more glucose than do normal cells from outside. Otto Heinrich Warburg (; 8 October 1883 – 1 August 1970) In 1931 was awarded the Nobel Prize in Physiology for his "discovery of the nature and mode of action of the respiratory enzyme. WARNBURG EFFECT : cancer cells under aerobic (well-oxygenated) conditions to metabolize glucose to lactate (aerobic glycolysis) is known as the Warburg effect. Warburg made the observation that tumor slices consume glucose and secrete lactate at a higher rate than normal tissues.

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PESTS OF SUGARCANE - BORERS BORERS 1. Early shoot borer, Chilo infuscatellus, Crambidae, Lepidoptera Symptom of damage: Deadheart in 1-3 month old crop, which can be easily pulled out, rotten portion of the straw coloured dead-heart emits an offensive odour. A number of bore holes at the base of the shoot just above the ground level. Nature of damage: Caterpillar bores into the central shoot and feeds on the internal tissue. This cause interference in the translocation of plant sap and damage the sugar storing tissues. Egg: Flat-scale like eggs are laid in 3-5 rows on the lower surface of leaves in masses of 4-100. The masses are slightly overlapping like tiles. Larva: Larva is dirty white with five dark violet longitudinal stripes and dark brown head. Pupa: Pupation takes place within the tunnel. Caterpillar before pupating makes a large exit hole in the stem and blocks the opening with silken discs. Adult: Pale greyish brown moth with black dots near the costal margin of the forewings and with white hindwings. 2. Internode borer, Chilo sacchariphagus indicus, Crambidae, Lepidoptera Symptom of damage: Internodes constricted and shortened, with a number of boreholes and fresh excreta in the nodal region. Affected tissues reddened. Nature of damage: Caterpillars attack sugarcane plants after 3 months of planting. They bore into the canes near the nodes; entry holes are plugged with excreta. Entry is generally confined to the first five internodes. Egg: Scale-like white eggs are laid by female moths in batches of 9-11, near the midribs, on leaf sheaths or on stem. Larva: White larva with four violet longitudinal stripes and light brown head. Pupa: Pupation takes place in semi-dried leaf sheath. Adult: Pale brown with white hind wings. 3. Top borer, Scirpophaga excerptalis, Pyralidae, Lepidoptera Symptom of damage: Deadheart in grownup canes, which cannot be easily pulled; deadheart reddish brown in colour; parallel row of shot holes in the emerging leaves and red tunnels in the midribs of leaves; bunchy tip due to the growth of side shoots. Nature of damage: Caterpillars are mainly found in the apical portion of the canes, boring through the growing point and down the upper joints until it reaches the sappy portion of the stem, there it feeds on the tissues and destroys the cane. They also bore into the unfolded leaves preferably into the midrib, mining its way to the base. Egg: Eggs are laid on the lower surface of top leaves in clusters particularly near midribs. The clusters are covered with buff coloured hairs. Larva :Smooth, white or cream coloured with a red coloured mid-dorsal line and yellow head. Pupa: Pupation takes place within the larval tunnel in a chamber with an exit hole constructed by the caterpillar. Adult: White coloured moth (with a buff coloured anal tuft in the abdominal tip of female).

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Introduction: RNA interference (RNAi) or Post-Transcriptional Gene Silencing (PTGS) is an important biological process for modulating eukaryotic gene expression. It is highly conserved process of posttranscriptional gene silencing by which double stranded RNA (dsRNA) causes sequence-specific degradation of mRNA sequences. dsRNA-induced gene silencing (RNAi) is reported in a wide range of eukaryotes ranging from worms, insects, mammals and plants. This process mediates resistance to both endogenous parasitic and exogenous pathogenic nucleic acids, and regulates the expression of protein-coding genes. What are small ncRNAs? micro RNA (miRNA) short interfering RNA (siRNA) Properties of small non-coding RNA: Involved in silencing mRNA transcripts. Called “small” because they are usually only about 21-24 nucleotides long. Synthesized by first cutting up longer precursor sequences (like the 61nt one that Lee discovered). Silence an mRNA by base pairing with some sequence on the mRNA. Discovery of siRNA? The first small RNA: In 1993 Rosalind Lee (Victor Ambros lab) was studying a non- coding gene in C. elegans, lin-4, that was involved in silencing of another gene, lin-14, at the appropriate time in the development of the worm C. elegans. Two small transcripts of lin-4 (22nt and 61nt) were found to be complementary to a sequence in the 3' UTR of lin-14. Because lin-4 encoded no protein, she deduced that it must be these transcripts that are causing the silencing by RNA-RNA interactions. Types of RNAi ( non coding RNA) MiRNA Length (23-25 nt) Trans acting Binds with target MRNA in mismatch Translation inhibition Si RNA Length 21 nt. Cis acting Bind with target Mrna in perfect complementary sequence Piwi-RNA Length ; 25 to 36 nt. Expressed in Germ Cells Regulates trnasposomes activity MECHANISM OF RNAI: First the double-stranded RNA teams up with a protein complex named Dicer, which cuts the long RNA into short pieces. Then another protein complex called RISC (RNA-induced silencing complex) discards one of the two RNA strands. The RISC-docked, single-stranded RNA then pairs with the homologous mRNA and destroys it. THE RISC COMPLEX: RISC is large(>500kD) RNA multi- protein Binding complex which triggers MRNA degradation in response to MRNA Unwinding of double stranded Si RNA by ATP independent Helicase Active component of RISC is Ago proteins( ENDONUCLEASE) which cleave target MRNA. DICER: endonuclease (RNase Family III) Argonaute: Central Component of the RNA-Induced Silencing Complex (RISC) One strand of the dsRNA produced by Dicer is retained in the RISC complex in association with Argonaute ARGONAUTE PROTEIN : 1.PAZ(PIWI/Argonaute/ Zwille)- Recognition of target MRNA 2.PIWI (p-element induced wimpy Testis)- breaks Phosphodiester bond of mRNA.)RNAse H activity. MiRNA: The Double-stranded RNAs are naturally produced in eukaryotic cells during development, and they have a key role in regulating gene expression .

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Cunningham, Fisiologia veterinaria, cuarta edición

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