Hershey and Chase Experiment

The Hershey–Chase experiments were a series of experiments conducted in 1952[1] by Alfred Hershey and Martha Chase that helped to confirm that DNA is the genetic material. While DNA had been known to biologists since 1869,[2] many scientists still assumed at the time that proteins carried the information for inheritance because DNA appeared simpler than proteins. In their experiments, Hershey and Chase showed that when bacteriophages, which are composed of DNA and protein, infect bacteria, their DNA enters the host bacterial cell, but most of their protein does not. Although the results were not conclusive, and Hershey and Chase were cautious in their interpretation, previous, contemporaneous and subsequent discoveries all served to prove that DNA is the hereditary material. Knowledge of DNA gained from these discoveries has applications in forensics, crime investigation and genealogy.[clarification needed]

Hershey shared the 1969 Nobel Prize in Physiology or Medicine with Max Delbrück and Salvador Luria for their “discoveries concerning the genetic structure of viruses

Horizontal gene transfer

Horizontal gene transfer (HGT) refers to the transfer of genes between organisms in a manner other than traditional reproduction. Also termed lateral gene transfer (LGT), it contrasts with vertical transfer, the transmission of genes from the parental generation to offspring via sexual or asexual reproduction. HGT has been shown to be an important factor in the evolution of many organisms.

Horizontal gene transfer is the primary reason for bacterial antibiotic resistance,and plays an important role in the evolution of bacteria that can degrade novel compounds such as human-created pesticides and in the evolution, maintenance, and transmission of virulence.This horizontal gene transfer often involves temperate bacteriophages and plasmids. Genes that are responsible for antibiotic resistance in one species of bacteria can be transferred to another species of bacteria through various mechanisms (e.g., via F-pilus), subsequently arming the antibiotic resistant genes' recipient against antibiotics, which is becoming a medical challenge to deal with.

Most thinking in genetics has focused upon vertical transfer, but there is a growing awareness that horizontal gene transfer is a highly significant phenomenon and among single-celled organisms perhaps the dominant form of genetic transfer.

Artificial horizontal gene transfer is a form of genetic engineering.

Hormonal Communication

Hormones are important messages both within the brain and between the brain and the body.

In addition to the nervous system, the endocrine system is a major communication system of the body. While the nervous system uses neurotransmitters as its chemical signals, the endocrine system uses hormones. The pancreas, kidneys, heart, adrenal glands, gonads, thyroid, parathyroid, thymus, and even fat are all sources of hormones. The endocrine system works in large part by acting on neurons in the brain, which controls the pituitary gland. The pituitary gland secretes factors into the blood that act on the endocrine glands to either increase or decrease hormone production. This is referred to as a feedback loop, and it involves communication from the brain to the pituitary to an endocrine gland and back to the brain. This system is very important for the activation and control of basic behavioral activities, such as sex; emotion; responses to stress; and eating, drinking, and the regulation of body functions, including growth, reproduction, energy use, and metabolism. The way the brain responds to hormones indicates that the brain is very malleable and capable of responding to environmental signals.

The brain contains receptors for thyroid hormones (those produced by the thyroid) and the six classes of steroid hormones, which are synthesized from cholesterol — androgens, estrogens, progestins, glucocorticoids, mineralocorticoids, and vitamin D. The receptors are found in selected populations of neurons in the brain and relevant organs in the body. Thyroid and steroid hormones bind to receptor proteins that in turn bind to DNA and regulate the action of genes. This can result in long-lasting changes in cellular structure and function.

The brain has receptors for many hormones; for example, the metabolic hormones insulin, insulin-like growth factor, ghrelin, and leptin. These hormones are taken up from the blood and act to affect neuronal activity and certain aspects of neuronal structure.

In response to stress and changes in our biological clocks, such as day and night cycles and jet lag, hormones enter the blood and travel to the brain and other organs. In the brain, hormones alter the production of gene products that participate in synaptic neurotransmission as well as affect the structure of brain cells. As a result, the circuitry of the brain and its capacity for neurotransmission are changed over a course of hours to days. In this way, the brain adjusts its performance and control of behavior in response to a changing environment.

Hormones are important agents of protection and adaptation, but stress and stress hormones, such as the glucocorticoid cortisol, can also alter brain function, including the brain’s capacity to learn. Severe and prolonged stress can impair the ability of the brain to function normally for a period of time, but the brain is also capable of remarkable recovery.

Reproduction in females is a good example of a regular, cyclic process driven by circulating hormones and involving a feedback loop: The neurons in the hypothalamus produce gonadotropin-releasing hormone (GnRH), a peptide that acts on cells in the pituitary. In both males and females, this causes two hormones — the follicle-stimulating hormone (FSH) and the luteinizing hormone (LH) — to be released into the bloodstream. In females, these hormones act on the ovary to stimulate ovulation and promote release of the ovarian hormones estradiol and progesterone. In males, these hormones are carried to receptors on cells in the testes, where they promote spermatogenesis and release the male hormone testosterone, an androgen, into the bloodstream. Testosterone, estrogen, and progesterone are often referred to as sex hormones.

In turn, the increased levels of testosterone in males and estrogen in females act on the hypothalamus and pituitary to decrease the release of FSH and LH. The increased levels of sex hormones also induce changes in cell structure and chemistry, leading to an increased capacity to engage in sexual behavior. Sex hormones also exert widespread effects on many other functions of the brain, such as attention, motor control, pain, mood, and memory.

Sexual differentiation of the brain is caused by sex hormones acting in fetal and early postnatal life, although recent evidence suggests genes on either the X or Y chromosome may also contribute to this process. Scientists have found statistically and biologically significant differences between the brains of men and women that are similar to sex differences found in experimental animals. These include differences in the size and shape of brain structures in the hypothalamus and the arrangement of neurons in the cortex and hippocampus. Sex differences go well beyond sexual behavior and reproduction and affect many brain regions and functions, ranging from mechanisms for perceiving pain and dealing with stress to strategies for solving cognitive problems. That said, however, the brains of men and women are more similar than they are different.

Anatomical differences have also been reported between the brains of heterosexual and homosexual men. Research suggests that hormones and genes act early in life to shape the brain in terms of sex-related differences in structure and function, but scientists are still putting together all the pieces of this puzzle.

How LASIK Surgery is done

LASIK, or "laser-assisted in situ keratomileusis," is the most commonly performed laser eye surgery to treat myopia (nearsightedness), hyperopia(farsightedness) and astigmatism.

Like other types of refractive surgery, the LASIK procedure reshapes the cornea to enable light entering the eye to be properly focused onto the retina for clearer vision.
In most cases, laser eye surgery is pain-free and completed within 15 minutes for both eyes. The results — improved vision without eyeglasses or contact lenses — can usually be seen in as little as 24 hours.
If you're not a good LASIK candidate, a number of other vision correction surgeries are available, such as PRK and LASEK laser eye surgery and phakic IOL surgery. Your eye doctor will determine if one of these procedures is suitable for your condition and, if so, which technique is best.

Want a visual? View our LASIK slide show!

How Is LASIK Surgery Performed?

First, your eye surgeon uses either a mechanical surgical tool called a microkeratome or a femtosecond laser to create a thin, circular "flap" in the cornea.
The surgeon then folds back the hinged flap to access the underlying cornea (called the stroma) and removes some corneal tissue using an excimer laser.
This highly specialized laser uses a cool ultraviolet light beam to remove ("ablate") microscopic amounts of tissue from the cornea to reshape it so it more accurately focuses light on the retina for improved vision.
For nearsighted people, the goal is to flatten the cornea; with farsighted people, a steeper cornea is desired.

Excimer lasers also can correct astigmatism by smoothing an irregular cornea into a more normal shape. It is a misconception that LASIK cannot treat astigmatism.

How Lungs Work

Why are the lungs important?
Oxygen, a basic gas, is needed by every cell in your body in order to live. The air that comes into the body through the lungs contains oxygen and other gases. In the lungs, the oxygen is moved into the bloodstream and carried through the body. At each cell in the body, the oxygen cells are exchanged for waste gas called carbon dioxide. The bloodstream then carries this waste gas back to the lungs where the waste gas is removed from the blood stream and then exhaled from the body. This vital process, called gas exchange, is performed automatically by the lungs and respiratory system.

In addition to gas exchange, the respiratory system performs other roles important to breathing. These include:

• Bringing air to the proper body temperature.
• Moisturizing the inhaled air to the right humidity.
• Protecting the body from harmful substances. This is done by coughing, sneezing, filtering, or swallowing them.
• The sense of smell.

The Parts of the Respiratory System and How They Work

AIRWAYS
The SINUSES are hollow spaces in the bones of the head. Small openings connect them to the nose. The functions they serve include helping to regulate the temperature and humidity of air breathed in.

The NOSE is the preferred entrance for outside air into the respiratory system. The hairs that line the wall are part of the air-cleaning system.

Air also enters through the MOUTH, especially in people who have a mouth-breathing habit or whose nasal passages may be temporarily obstructed, as by a cold or during heavy exercise.

The THROAT collects incoming air from the nose and mouth and passes it downward to the windpipe (trachea).

The WINDPIPE (trachea) is the passage leading from the throat to the lungs.

The windpipe divides into the two main BRONCHIAL TUBES, one for each lung, which subdivide into each lobe of the lungs. These, in turn, subdivide further.

Lungs and Blood Vessels
The right lung is divided into three LOBES, or sections. Each lobe is like a balloon filled with sponge-like tissue. Air moves in and out through one opening -- a branch of the bronchial tube.

The left lung is divided into two LOBES.

The PLEURA are the two membranes, actually one continuous one folded on itself, that surround each lobe of the lungs and separate the lungs from the chest wall.

The bronchial tubes are lined with CILIA (like very small hairs) that have a wave-like motion. This motion carries MUCUS (sticky phlegm or liquid) upward and out into the throat, where it is either coughed up or swallowed. The mucus catches and holds much of the dust, germs, and other unwanted matter that has invaded the lungs. You get rid of this matter when you cough, sneeze, clear your throat or swallow.

The smallest subdivisions of the bronchial tubes are called BRONCHIOLES, at the end of which are the air sacs or alveoli.

The ALVEOLI are the very small air sacs that are the destination of air breathed in.

The CAPILLARIES are blood vessels that are imbedded in the walls of the alveoli. Blood passes through the capillaries, brought to them by the PULMONARY ARTERY and taken away by the PULMONARY VEIN. While in the capillaries the blood gives off carbon dioxide through the capillary wall into the alveoli and takes up oxygen from the air in the alveoli.

Muscles and Bones
The DIAPHRAGM is the strong wall of muscle that separates the chest cavity from the abdominal cavity. By moving downward, it creates suction in the chest to draw in air and expand the lungs.

The RIBS are bones supporting and protecting the chest cavity. They move to a limited degree, helping the lungs to expand and contract.

Keeping Lungs Healthy

Taking good care of yourself every day will help keep your lungs healthy. Good health habits like eating a balanced diet, exercising and reducing the stress in your life will help you breathe easier.

How Nucleotides are added in DNA Replication

DNA replication is the process of producing two identical replicas from one original DNA molecule. This biological process occurs in all living organisms and is the basis for biological inheritance. DNA is made up of two strands and each strand of the original DNA molecule serves as template for the production of the complementary strand, a process referred to as semiconservative replication. Cellular proofreading and error-checking mechanisms ensure near perfect fidelity for DNA replication.

In a cell, DNA replication begins at specific locations, or origins of replication, in the genome.Unwinding of DNA at the origin and synthesis of new strands results in replication forks growing bidirectional from the origin. A number of proteins are associated with the replication fork which helps in terms of the initiation and continuation of DNA synthesis. Most prominently, DNA polymerase synthesizes the new DNA by adding complementary nucleotides to the template strand.

DNA replication can also be performed in vitro (artificially, outside a cell). DNA polymerases isolated from cells and artificial DNA primers can be used to initiate DNA synthesis at known sequences in a template DNA molecule. The polymerase chain reaction (PCR), a common laboratory technique, cyclically applies such artificial synthesis to amplify a specific target DNA fragment from a pool of DNA.

How Spliceosomes Process RNA

A spliceosome is a large and complex molecular machine found primarily within the splicing speckles of the cell nucleus of eukaryotic cells. The spliceosome is assembled from snRNPs and protein complexes. The spliceosome removes introns from a transcribed pre-mRNA, a kind of primary transcript. This process is generally referred to as splicing. Only eukaryotes have spliceosomes and metazoans have a second spliceosome, the minor spliceosome.

How the Krebs Cycle Works

The citric acid cycle – also known as the tricarboxylic acid cycle (TCA cycle), or the Krebs cycle, – is a series of chemical reactions used by all aerobic organisms to generate energy through the oxidation of acetate derived from carbohydrates, fats and proteins into carbon dioxide and chemical energy in the form of adenosine triphosphate (ATP). In addition, the cycle provides precursors of certain amino acids as well as the reducing agent NADH that is used in numerous other biochemical reactions. Its central importance to many biochemical pathways suggests that it was one of the earliest established components of cellular metabolism and may have originated abiogenically.
The name of this metabolic pathway is derived from citric acid (a type of tricarboxylic acid) that is consumed and then regenerated by this sequence of reactions to complete the cycle. In addition, the cycle consumes acetate (in the form of acetyl-CoA) and water, reduces NAD+ to NADH, and produces carbon dioxide as a waste byproduct. The NADH generated by the TCA cycle is fed into the oxidative phosphorylation (electron transport) pathway. The net result of these two closely linked pathways is the oxidation of nutrients to produce usable chemical energy in the form of ATP.

In eukaryotic cells, the citric acid cycle occurs in the matrix of the mitochondrion. In prokaryotic cells, such as bacteria which lack mitochondria, the TCA reaction sequence is performed in the cytosol with the proton gradient for ATP production being across the cell's surface (plasma membrane) rather than the inner membrane of the mitochondrion.

How the NAD+ Works

How the NAD+ Works
Nicotinamide adenine dinucleotide, abbreviated NAD+, is a coenzyme found in all living cells. The compound is a dinucleotide, since it consists of two nucleotides joined through their phosphate groups: with one nucleotide containing an adenosine ring, and the other containing nicotinamide.
In metabolism, NAD+ is involved in redox reactions, carrying electrons from one reaction to another. The coenzyme is therefore found in two forms in cells: NAD+ is an oxidizing agent – it accepts electrons from other molecules and becomes reduced, this reaction forms NADH, which can then be used as a reducing agent to donate electrons. These electron transfer reactions are the main function of NAD+. However, it is also used in other cellular processes, notably as a substrate of enzymes that add or remove chemical groups from proteins, in posttranslational modifications. Due to the importance of these functions, the enzymes involved in NAD+ metabolism are targets for drug discovery.

In organisms, NAD+ can be synthesized from scratch (de novo) from the amino acids tryptophan or aspartic acid. Alternatively, components of the coenzymes are taken up from food as the vitamin called niacin. Similar compounds are released by reactions that break down the structure of NAD+. These preformed components then pass through a salvage pathway that recycles them back into the active form. Some NAD+ is also converted into nicotinamide adenine dinucleotide phosphate (NADP+); the chemistry of this related coenzyme is similar to that of NAD+, but it has different roles in metabolism.

Nicotinamide adenine dinucleotide is a dinucleotide since it consists of two nucleotides joined by a pair of bridging phosphate groups. The nucleotides consist of ribose rings, one with adenine attached to the first carbon atom (the 1' position) and the other with nicotinamide at this position. The nicotinamide group can be attached in two orientations to this anomeric carbon atom, due to these two possible structures, the compound exists as two diastereomers. It is the β-nicotinamide diastereomer of NAD+ which is found in organisms. These nucleotides are joined together by a bridge of two phosphate groups through the 5' carbons. In metabolism the compound accepts or donates electrons in redox reactions.Such reactions (summarized in formula below) involve the removal of two hydrogen atoms from the reactant (R), in the form of a hydride ion, and a proton (H+). The proton is released into solution, while the reductant RH2 is oxidized and NAD+ reduced to NADH by transfer of the hydride to the nicotinamide ring. RH2 + NAD+ → NADH + H+ + R From the hydride electron pair, one electron is transferred to the positively-charged nitrogen of the nicotinamide ring of NAD+, and the second hydrogen atom transferred to the C4 carbon atom opposite this nitrogen. The midpoint potential of the NAD+/NADH redox pair is −0.32 volts, which makes NADH a strong reducing agent. The reaction is easily reversible, when NADH reduces another molecule and is re-oxidized to NAD+. This means the coenzyme can continuously cycle between the NAD+ and NADH forms without being consumed. In appearance, all forms of this coenzyme are white amorphous powders that are hygroscopic and highly water-soluble. The solids are stable if stored dry and in the dark. Solutions of NAD+ are colorless and stable for about a week at 4 °C and neutral pH, but decompose rapidly in acids or alkalis. Upon decomposition, they form products that are enzyme inhibitors. Both NAD+ and NADH absorb strongly in the ultraviolet due to the adenine base. For example, peak absorption of NAD+ is at a wavelength of 259 nanometers (nm), with an extinction coefficient of 16,900 M-1cm-1. NADH also absorbs at higher wavelengths, with a second peak in UV absorption at 339 nm with an extinction coefficient of 6,220 M-1cm-1. This difference in the ultraviolet absorption spectra between the oxidized and reduced forms of the coenzymes at higher wavelengths makes it simple to measure the conversion of one to another in enzyme assays – by measuring the amount of UV absorption at 340 nm using a spectrophotometer. NAD+ and NADH also differ in their fluorescence. NADH in solution has an emission peak at 460 nm and a fluorescence lifetime of 0.4 nanoseconds, while the oxidized form of the coenzyme does not fluoresce. The properties of the fluorescence signal changes when NADH binds to proteins, so these changes can be used to measure dissociation constants, which are useful in the study of enzyme kinetics. These changes in fluorescence are also used to measure changes in the redox state of living cells, through fluorescence microscopy.

How Translation Works

Integration and Excision of a Plasmid

Multicopy plasmids have greatly facilitated gene structure-function studies. However, the use of such plasmids can lead to high-copy-number artifacts, especially in physiological studies. Thus, several methods have been developed for recombining genes on bacterial chromosomes in order to study their functions in single copies. Such methods are frequently used to construct novel Escherichia coli strains that stably express foreign genes for use in both basic research and biotechnology (5, 18, 27). However, the development of strains encoding complex metabolic or regulatory pathways poses special problems that often require manipulating many genes and expressing them individually at different levels or under separate regulatory controls. To address these concerns, we have developed a series of plasmid-host systems for the introduction of multiple genes into the same cell in single copies. Our approach is based on genome targeting systems that utilize plasmids carrying a conditional-replication origin and a phage attachment (attP) site (17). We refer to our plasmids as CRIM (conditional-replication, integration, and modular) plasmids. CRIM plasmids can be integrated into or retrieved from their bacterial attachment (attB) site by supplying phage integrase (Int) without or with excisionase (Xis) in trans.

Advantages of our CRIM plasmid-host systems include the use of alternative attP andattB sites (for phages λ, HK022, 80, P21, and P22) and different selectable markers (for chloramphenicol, gentamicin, kanamycin, spectinomycin and streptomycin, tetracycline, and trimethoprim resistance) in conjunction with a polylinker or promoter (ParaB, PrhaB,PrhaS, Ptac, Psyn1, and Psyn4) for ectopic expression of the cloned gene(s). These CRIM plasmids have the γ replication origin of R6K, which requires the trans-acting Π protein (encoded by pir) for replication. So, they replicate at a medium (15 per cell) or high (250 per cell) plasmid copy number in pir+ or pir-116 (high-copy-number mutant) E. coli hosts (28), respectively. Int helper plasmids are used for integration of CRIM plasmids into the corresponding chromosomal attB sites of normal (non-pir) hosts, which are nonpermissive for CRIM plasmid replication. Xis/Int helper plasmids are used for excision (“curing”) of the respective CRIM plasmids from the chromosome, e.g., to verify that phenotypes are due to their presence. Xis/Int helper plasmids are also used for retrieval (cloning) of CRIM plasmids from the chromosome, e.g., to recover a particular CRIM plasmid after screening of CRIM plasmid or mutant libraries.

Since integration and retrieval involve phage-site-specific recombination events, the original and recovered plasmids are identical. CRIM plasmids can therefore be used for the construction of gene (or mutant) libraries that can be directly integrated into bacterial chromosomes in single copies for screening or selection purposes. Afterwards, CRIM plasmids can be retrieved from individual cells or en masse. The recovered plasmids can then be propagated as plasmids for molecular analysis or integrated directly into the chromosomes of other hosts for subsequent processing without further in vitro manipulation steps. We previously found similar oriRγ attλ plasmids to be extremely useful in mutagenesis studies, especially when it was important that the mutated gene be free of plasmid copy number effects (16). We also found them to be useful in studying genes from diverse bacteria, including gram-negative and -positive cells (14, 15, 25, 34). Our versatile CRIM plasmid-host systems should be widely useful in gene structure-function studies. Here we describe our basic set of CRIM plasmids, the requisite helper plasmids, and how to use them.

Ionic vs Covalent Bonding

Covalent Bonds vs Ionic Bonds

There are two types of atomic bonds - ionic bonds and covalent bonds. They differ in their structure and properties. Covalent bondsconsist of pairs of electrons shared by two atoms, and bind the atoms in a fixed orientation. Relatively high energies are required to break them (50 - 200 kcal/mol). Whether two atoms can form a covalent bond depends upon their electronegativity i.e. the power of an atom in a molecule to attract electrons to itself. If two atoms differ considerably in their electronegativity - as sodium and chloride do - then one of the atoms will lose its electron to the other atom. This results in a positively charged ion (cation) and negatively charged ion (anion). The bond between these two ions is called an ionic bond.

	Covalent Bonds	Ionic Bonds
State at room temperature:	Liquid or gaseous	Solid
Polarity:	Low	High
Formation:	A covalent bond is formed between two non-metals that have similar electronegativities. Neither atom is "strong" enough to attract electrons from the other. For stabilization, they share their electrons from outer molecular orbit with others	An ionic bond is formed between a metal and a non-metal. Non-metals(-ve ion) are "stronger" than the metal(+ve ion) and can get electrons very easily from the metal. These two opposite ions attract each other and form the ionic bond.
Shape:	Definite shape	No definite shape
Melting point:	low	High
What is it?:	Covalent bonding is a form of chemical bonding between two non metallic atoms which is characterized by thesharing of pairs of electrons between atoms and other covalent bonds.	Ionic bond, also known as electrovalent bond, is a type of bond formed from the electrostatic attraction between oppositely charged ions in a chemical compound. These kinds of bonds occur mainly between a metallic and a non metallic atom.
Boiling point:	Low	High
Examples:	Methane (CH4), Hydro Chloric acid (HCl)	Sodium chloride (NaCl), Sulphuric Acid (H2SO4 )
Occurs between:	Two non-metals

Kidney Function Counter current Mechanism

A countercurrent multiplier system is a mechanism that expends energy to create a concentration gradient.

It is found widely in nature and especially in mammalian organs. For example, it can refer to the process that is underlying the process of urine concentration, that is, the production of hyperosmotic urine by the mammalian kidney. The ability to concentrate urine is also present in birds.[

Countercurrent multiplication is frequently mistaken for countercurrent exchange, a similar but different mechanism where gradients are maintained, but not established.

Physiological principles
The term derives from the form and function of the loop of Henle, which consists of two parallel limbs of renal tubules running in opposite directions, separated by the interstital space of the renal medulla.

The descending limb of the loop of Henle is permeable to water but impermeable to solutes, due to the presence of aquaporin 1 in its tubular wall. Thus water moves across the tubular wall into the medullary space, making the filtrate hypertonic.
The ascending limb is impermeable to water (because of a lack of aquaporin, a common transporter protein for water channels in all cells except the walls of the ascending limb of the loop of Henle) but permeable to solutes, but here Na+, Cl−, and K+ are actively transported into the medullary space, making the filtrate hypotonic (with a higher water potential). This constitutes the single effect of the countercurrent multiplication process.
Active transport of these ions from the thick ascending limb creates an osmotic pressure drawing water from the descending limb into the hyperosmolar medullary space, making the filtrate hypertonic (with a lower water potential).
The countercurrent flow within the descending and ascending limb thus increases, or multiplies the osmotic gradient between tubular fluid and interstitial space.

Details
Countercurrent multiplication was originally studied as a mechanism whereby urine is concentrated in the nephron. Initially studied in the 1950s by Gottschalk and Mylle following Werner Kuhn's postulations,this mechanism gained popularity only after a series of complicated micropuncture experiments.

The proposed mechanism consists of pump, equilibration, and shift steps. In the proximal tubule, the osmolarity is isomolar to plasma (300 mOsm/L). In a hypothetical model where there was no equilibration or pump steps, the tubular fluid and interstitial osmolarity would be 300 mOsm/L as well.{Respicius Rwehumbiza, 2010}

Pump: The Na+/K+/2Cl− transporter in the ascending limb of the loop of Henle helps to create a gradient by shifting Na+ into the medullary interstitium. The thick ascending limb of the loop of Henle is the only part of the nephron lacking in aquaporin—a common transporter protein for water channels. This makes the thick ascending limb impermeable to water. The action of the Na+/K+/2Cl− transporter therefore creates a hypoosmolar solution in the tubular fluid and a hyperosmolar fluid in the interstitium, since water cannot follow the solutes to produce osmotic equilibrium.

Equilibration: Since the descending limb of the loop of henle consists of very leaky epithelium, the fluid inside the descending limb becomes hyperosmolar.

Shift: The movement of fluid through the tubules causes the hyperosmotic fluid to move further down the loop. Repeating many cycles causes fluid to be near isosmolar at the top of Henle's loop and very concentrated at the bottom of the loop. Interestingly, animals with a need for very concentrated urine (such as desert animals) have very long loops of Henle to create a very large osmotic gradient. Animals that have abundant water on the other hand (such as beavers) have very short loops. The vasa recta have a similar loop shape so that the gradient does not dissipate into the plasma.

The mechanism of counter current multiplication works together with the vasa recta's counter current exchange to prevent the wash out of salts and maintain a high osmolarity at the inner medulla

Kidney Function II

The kidneys are bean-shaped organs that serve several essential regulatory roles in vertebrate animals. They remove excess organic molecules (e.g., glucose) from the blood, and it is by this action that their best-known function is performed: the removal of waste products of metabolism (e.g., urea, though 90% of this is reabsorbed along the nephron.) They are essential in the urinary system and also serve homeostatic functions such as the regulation of electrolytes, maintenance of acid–base balance, and regulation of blood pressure (via maintaining salt and water balance). They serve the body as a natural filter of the blood, and remove water soluble wastes, which are diverted to the urinary bladder. In producing urine, the kidneys excrete wastes such as urea and ammonium, and they are also responsible for the reabsorption of water, glucose, and amino acids. The kidneys also produce hormones including calcitriol, erythropoietin, and the enzyme renin, the last of which indirectly acts on the kidney in negative feedback.

Located at the rear of the abdominal cavity in the retroperitoneum, the kidneys receive blood from the paired renal arteries, and drain into the paired renal veins. Each kidney excretes urine into a ureter, itself a paired structure that empties into the urinary bladder.

Renal physiology is the study of kidney function, while nephrology is the medical specialty concerned with kidney diseases. Diseases of the kidney are diverse, but individuals with kidney disease frequently display characteristic clinical features. Common clinical conditions involving the kidney include the nephritic and nephrotic syndromes, renal cysts, acute kidney injury, chronic kidney disease, urinary tract infection, nephrolithiasis, and urinary tract obstruction. Various cancers of the kidney exist; the most common adult renal cancer is renal cell carcinoma. Cancers, cysts, and some other renal conditions can be managed with removal of the kidney, or nephrectomy. When renal function, measured by glomerular filtration rate, is persistently poor, dialysis and kidney transplantation may be treatment options. Although they are not normally harmful, kidney stones can be painful, and repeated, chronic formation of stones can scar the kidneys. The removal of kidney stones involves ultrasound treatment to break up the stones into smaller pieces, which are then passed through the urinary tract. One common symptom of kidney stones is a sharp to disabling pain in the medial/lateral segments of the lower back or groin.

Lymphatic System

The lymphatic system is part of the circulatory system, comprising a network of lymphatic vessels that carry a clear fluid called lymph (from Latin lympha meaning water) directionally towards the heart. The lymphatic system was first described in the seventeenth century independently by Olaus Rudbeck and Thomas Bartholin. Unlike the cardiovascular system the lymphatic system is not a closed system. The human circulatory system processes an average of 20 litres of blood per day through capillary filtration which removes plasma while leaving the blood cells. Roughly 17 litres of the filtered plasma get reabsorbed directly into the blood vessels, while the remaining 3 litres are left behind in the interstitial fluid. One of the main functions of the lymph system is to provide an accessory route for these excess 3 litres per day to get returned to the blood.

The other main function is that of defense in the immune system. Lymph is very similar to blood plasma but contains lymphocytes and other white blood cells. It also contains waste products and debris of cells together with bacteria and protein. Associated organs composed of lymphoid tissue are the sites of lymphocyte production. Lymphocytes are concentrated in the lymph nodes. The spleen and the thymus are also lymphoid organs of the immune system. The tonsils are lymphoid organs that are also associated with the digestive system. Lymphoid tissues contain lymphocytes, and also contain other types of cells for support. The system also includes all the structures dedicated to the circulation and production of lymphocytes (the primary cellular component of lymph), which also includes the bone marrow, and the lymphoid tissue associated with the digestive system.

The blood does not come into direct contact with the parenchymal cells and tissues in the body, but constituents of the blood first exit the microvascular exchange blood vessels to become interstitial fluid, which comes into contact with the parenchymal cells of the body. Lymph is the fluid that is formed when interstitial fluid enters the initial lymphatic vessels of the lymphatic system. The lymph is then moved along the lymphatic vessel network by either intrinsic contractions of the lymphatic passages or by extrinsic compression of the lymphatic vessels via external tissue forces (e.g. the contractions of skeletal muscles), or by lymph hearts in some animals. The organization of lymph nodes and drainage follows the organization of the body into external and internal regions; therefore, the lymphatic drainage of the head, limbs, and body cavity walls follows an external route, and the lymphatic drainage of the thorax, abdomen, and pelvic cavities follows an internal route.Eventually, the lymph vessels empty into the lymphatic ducts, which drain into one of the two subclavian veins, near their junction with the internal jugular veins.

Maturation of the Follicle

In biology, folliculogenesis is the maturation of the ovarian follicle, a densely packed shell of somatic cells that contains an immature oocyte. Folliculogenesis describes the progression of a number of small primordial follicles into large preovulatory follicles that enter the menstrual cycle.

Contrary to male spermatogenesis, which can last indefinitely, folliculogenesis ends when the remaining follicles in the ovaries are incapable of responding to the hormonal cues that previously recruited some follicles to mature. This depletion in follicle supply signals the beginning of menopause.

Meselson and Stahl Experiment

The Meselson–Stahl experiment was an experiment by Matthew Meselson and Franklin Stahl in 1958 which supported the hypothesis that DNA replication was semiconservative. In semiconservative replication, when the double stranded DNA helix is replicated each of the two new double-stranded DNA helixes consisted of one strand from the original helix and one newly synthesized. It has been called "the most beautiful experiment in biology

Hypothesis

A summary of the three postulated methods of DNA synthesis
Three hypotheses had been previously proposed for the method of replication of DNA.

In the semiconservative hypothesis, proposed by Watson and Crick, the two strands of a DNA molecule separate during replication. Each strand then acts as a template for synthesis of a new strand.

The conservative hypothesis proposed that the entire DNA molecule acted as a template for the synthesis of an entirely new one. According to this model, histone proteins bind to the DNA, revolving the strand and exposing the nucleotide bases (which normally line the interior) for hydrogen bonding.

The dispersive hypothesis is exemplified by a model proposed by Max Delbrück, which attempts to solve the problem of unwinding the two strands of the double helix by a mechanism that breaks the DNA backbone every 10 nucleotides or so, untwists the molecule, and attaches the old strand to the end of the newly synthesized one. This would synthesize the DNA in short pieces alternating from one strand to the other.

Each of these three models makes a different prediction about the distribution of the "old" DNA in molecules formed after replication. In the conservative hypothesis, after replication, one molecule is the entirely conserved "old" molecule, and the other is all newly synthesized DNA. The semiconservative hypothesis predicts that each molecule after replication will contain one old and one new strand. The dispersive model predicts that each strand of each new molecule will contain a mixture of old and new DNA.

Experimental procedure and results
Meselson-stahl experiment diagram en.svg
Nitrogen is a major constituent of DNA. 14N is by far the most abundant isotope of nitrogen, but DNA with the heavier (but non-radioactive) 15N isotope is also functional.

E. coli were grown for several generations in a medium with 15N. When DNA is extracted from these cells and centrifuged on a salt density gradient, the DNA separates out at the point at which its density equals that of the salt solution. The DNA of the cells grown in 15N medium had a higher density than cells grown in normal 14N medium. After that, E. coli cells with only 15N in their DNA were transferred to a 14N medium and were allowed to divide; the progress of cell division was monitored by microscopic cell counts and by colony assay.

DNA was extracted periodically and was compared to pure 14N DNA and 15N DNA. After one replication, the DNA was found to have intermediate density. Since conservative replication would result in equal amounts of DNA of the higher and lower densities (but no DNA of an intermediate density), conservative replication was excluded. However, this result was consistent with both semiconservative and dispersive replication. Semiconservative replication would result in double-stranded DNA with one strand of 15N DNA, and one of 14N DNA, while dispersive replication would result in double-stranded DNA with both strands having mixtures of 15N and 14N DNA, either of which would have appeared as DNA of an intermediate density.

The authors continued to sample cells as replication continued. DNA from cells after two replications had been completed was found to consist of equal amounts of DNA with two different densities, one corresponding to the intermediate density of DNA of cells grown for only one division in 14N medium, the other corresponding to DNA from cells grown exclusively in 14N medium. This was inconsistent with dispersive replication, which would have resulted in a single density, lower than the intermediate density of the one-generation cells, but still higher than cells grown only in 14N DNA medium, as the original 15N DNA would have been split evenly among all DNA strands. The result was consistent with the semiconservative replication hypothesis

Methyl directed mismatch repair

DNA mismatch repair is a system for recognizing and repairing erroneous insertion, deletion, and mis-incorporation of bases that can arise during DNA replication and recombination, as well as repairing some forms of DNA damage.

Mismatch repair is strand-specific. During DNA synthesis the newly synthesised (daughter) strand will commonly include errors. In order to begin repair, the mismatch repair machinery distinguishes the newly synthesised strand from the template (parental). In gram-negative bacteria, transient hemimethylation distinguishes the strands (the parental is methylated and daughter is not). However, in other prokaryotes and eukaryotes, the exact mechanism is not clear. It is suspected that, in eukaryotes, newly synthesized lagging-strand DNA transiently contains nicks (before being sealed by DNA ligase) and provides a signal that directs mismatch proofreading systems to the appropriate strand. This implies that these nicks must be present in the leading strand, and evidence for this has recently been found.[3] Recent work [4] has shown that nicks are sites for RFC-dependent loading of the replication sliding clamp PCNA, in an orientation-specific manner, such that one face of the donut-shape protein is juxtaposed toward the 3'-OH end at the nick. Oriented PCNA then directs the action of the MutLalpha endonuclease to one strand in the presence of a mismatch and MutSalpha or MutSbeta.

Any mutational event that disrupts the superhelical structure of DNA carries with it the potential to compromise the genetic stability of a cell. The fact that the damage detection and repair systems are as complex as the replication machinery itself highlights the importance evolution has attached to DNA fidelity.

Examples of mismatched bases include a G/T or A/C pairing (see DNA repair). Mismatches are commonly due to tautomerization of bases during G2. The damage is repaired by recognition of the deformity caused by the mismatch, determining the template and non-template strand, and excising the wrongly incorporated base and replacing it with the correct nucleotide. The removal process involves more than just the mismatched nucleotide itself. A few or up to thousands of base pairs of the newly synthesized DNA strand can be removed.

DNA mismatch repair (MMR) is an evolutionarily conserved process that corrects mismatches generated during DNA replication and escape proofreading. MMR proteins also participate in many other DNA transactions, such that inactivation of MMR can have wide-ranging biological consequences, which can be either beneficial or detrimental. We begin this review by briefly considering the multiple functions of MMR proteins and the consequences of impaired function. We then focus on the biochemical mechanism of MMR replication errors. Emphasis is on structure-function studies of MMR proteins, on how mismatches are recognized, on the process by which the newly replicated strand is identified, and on excision of the replication error.

Microarrays DNA Chips

A DNA microarray (also commonly known as DNA chip or biochip) is a collection of microscopic DNA spots attached to a solid surface. Scientists use DNA microarrays to measure the expression levels of large numbers of genes simultaneously or to genotype multiple regions of a genome. Each DNA spot contains picomoles (10−12 moles) of a specific DNA sequence, known as probes (or reporters or oligos). These can be a short section of a gene or other DNA element that are used to hybridize a cDNA or cRNA (also called anti-sense RNA) sample (called target) under high-stringency conditions. Probe-target hybridization is usually detected and quantified by detection of fluorophore-, silver-, or chemiluminescence-labeled targets to determine relative abundance of nucleic acid sequences in the target.

Micturition Reflex

Urination is the release of urine from the urinary bladder through the urethra to the urinary meatus outside of the body. It is also known medically as micturition, voiding, uresis, or, rarely, emiction, and known colloquially by various names including tinkling, peeing, weeing, and pissing. In healthy humans (and many other animals) the process of urination is under voluntary control. In infants, some elderly individuals, and those with neurological injury, urination may occur as an involuntary reflex. In some animals, in addition to expelling waste material, urination can mark territory or express submissiveness. Physiologically, urination involves coordination between the central, autonomic, and somatic nervous systems. Brain centers that regulate urination include the pontine micturition center, periaqueductal gray, and the cerebral cortex.[citation needed] In male placental mammals, urine is ejected through the penis. In female placental mammals, urine is ejected through the vulva or pseudo-penis