TERM PAPER

OF

CONCEPTS IN BIOTECHNOLOGY

Topic:-Role of antibiotics in controlling diseases

Submitted To:-

Mr. Harsh

Submitted By:-

Vidhu Sharma

Roll.no.:-47

Reg. No.1040070123

Lovely Professional University

CONTENTS
Introdoction

History

Types of antibiotics

Role of antibiotics in controlling diseases

Mechanism of action

Four types of mechanisms

Applications

ANTIBIOTICS
Introduction

Staphylococcus aureus - Antibiotics test plate.

An antibiotic is a chemotherapeutic agent that inhibits or abolishes the growth

of micro-organisms, such as bacteria, fungi, or protozoans. The term originally
referred to any agent with biological activity against living organisms; however,
"antibiotic" now is used to refer to substances with anti-bacterial, anti-fungal, or
anti-parasitical activity. The first antibiotic compounds used in modern
medicine were produced and isolated from living organisms, such as the
penicillin class produced by fungi in the genus Penicillium, or streptomycin
from bacteria of the genus Streptomyces.

.
Points of attack on bacteria by antibiotics

Anti-bacterial antibiotics can be categorized based on their target

specificity: "narrow-spectrum" antibiotics target particular types of bacteria,
such as Gram-negative or Gram-positive bacteria, while broad-spectrum
antibiotics affect a wide range of bacteria.The effectiveness of individual
antibiotics varies with the location of the infection, the ability of the antibiotic
to reach the site of infection, and the ability of the microbe to inactivate or
excrete the antibiotic. Some anti-bacterial antibiotics destroy bacteria
(bactericidal), whereas others prevent bacteria from multiplying (bacteriostatic).

Oral antibiotics are simply ingested, while intravenous antibiotics are used in
more serious cases, such as deep-seated systemic infections. Antibiotics may
also sometimes be administered topically, as with eye drops or ointments.
History

Penicillin

Although potent antibiotic compounds for treatment of human diseases

caused by bacteria (such as tuberculosis, bubonic plague, or leprosy) were not
isolated and identified until the twentieth century, the first known use of
antibiotics was by the ancient Chinese over 2,500 years ago.Many other ancient
cultures, including the ancient Egyptians and ancient Greeks already used molds
and plants to treat infections, owing to the production of antibiotic substances
by these organisms. At that time, however, the compounds having antibiotic
activity and present in moulds or plants were unknown.

The antibiotic properties of Penicillium sp. were first described in France by

Ernest Duchesne in 1897. However, his work went by without much notice
from the scientific community until Alexander Flemig's discovery of Penicillim.

Antibiotic peptides are important effector molecules in host-parasite interactions

throughout the living world. In vertebrates, they function in first-line host
Types of Antibiotics

Penicillins

Penicillins are bactericidal, inhibiting formation of the cell wall. There

are four types of penicillins: the narrow-spectrum penicillin-G types, ampicillin
and its relatives, the penicillinase-resistants, and the extended spectrum
penicillins that are active against pseudomonas. Penicillin-G types are effective
against gram-positive strains of streptococci, staphylococci, and some gram-
negative bacteria such as meningococcus. Penicillin-G is used to treat such
diseases as syphilis, gonorrhea, meningitis, anthrax, and yaws. Ampicillin and
amoxicillin have a range of effectiveness similar to that of penicillin-G, with a
slightly broader spectrum, including some gram-negative bacteria. The
penicillinase-resistants are penicillins that combat bacteria that have developed
resistance to penicillin-G. The antipseudomonal penicillins are used against
infections caused by gram-negative Pseudomonas bacteria, a particular problem
in hospitals. They may be administered as a prophylactic in patients with
compromised immune systems, who are at risk from gram-negative infections.

Cephalosporin

Cephalosporins have a Β-lactam ring structure that interferes with

synthesis of the bacterial cell wall and so are bactericidal. Cephalosporins are
more effective than penicillin against gram-negative bacilli and equally
effective against gram-positive cocci. Cephalosporins may be used to treat
strains of meningitis and as a prophylactic for orthopedic, abdominal, and pelvic
surgery. Rare hypersensitive reactions from the cephalosporins include skin rash
and, less frequently, anaphylactic shock.
Aminoglycosides

Streptomycin is the oldest of the aminoglycosides. The

aminoglycosides inhibit bacterial protein synthesis in many gram-negative and
some gram-positive organisms. They are sometimes used in combination with
penicillin. The members of this group tend to be more toxic than other
antibiotics. Rare adverse effects associated with prolonged use of
aminoglycosides include damage to the vestibular region of the ear, hearing
loss, and kidney damage.

Tetracylines

Tetracyclines are bacteriostatic, inhibiting bacterial protein synthesis.

They are broad-spectrum antibiotics effective against strains of streptococci,
gram-negative bacilli, rickettsia (the bacteria that causes typhoid fever), and
spirochetes (the bacteria that causes syphilis). They are also used to treat
urinary-tract infections and bronchitis. Because of their wide range of
effectiveness, tetracyclines can sometimes upset the balance of resident bacteria
that are normally held in check by the body's immune system, leading to
secondary infections in the gastrointestinal tract and vagina, for example.
Tetracycline use is now limited because of the increase of resistant bacterial
strains.

Macrolides

The macrolides are bacteriostatic, binding with bacterial ribosomes to

inhibit protein synthesis. Erythromycin, one of the macrolides, is effective
against gram-positive cocci and is often used as a substitute for penicillin
against streptococcal and pneumococcal infections. Other uses for macrolides
include diphtheria and bacteremia. Side effects may include nausea, vomiting,
and diarrhea; infrequently, there may be temporary auditory impairment.

Sulfonamides

The sulfonamides are synthetic bacteriostatic, broad-spectrum

antibiotics, effective against most gram-positive and many gram-negative
bacteria. However, because many gram-negative bacteria have developed
resistance to the sulfonamides, these antibiotics are now used only in very
specific situations, including treatment of urinary-tract infection, against
meningococcal strains, and as a prophylactic for rheumatic fever. Side effects
may include disruption of the gastrointestinal tract and hypersensitivity.

ROLE OF ANTIBIOTICS IN CONTROLLING

DISEASES

Most antibiotics act by selectively interfering with the synthesis of one of the
large-molecule constituents of the cell—the cell wall or proteins or nucleic
acids. Some, however, act by disrupting the cell membrane (see Cell Death and
Growth Suppression below). Some important and clinically useful drugs
interfere with the synthesis of peptidoglycan, the most important component of
the cell wall. These drugs include the Β-lactam antibiotics, which are classified
according to chemical structure into penicillins, cephalosporins, and
carbapenems. All these antibiotics contain a Β-lactam ring as a critical part of
their chemical structure, and they inhibit synthesis of peptidoglycan, an
essential part of the cell wall. They do not interfere with the synthesis of other
intracellular components. The continuing buildup of materials inside the cell
exerts ever greater pressure on the membrane, which is no longer properly
supported by peptidoglycan. The membrane gives way, the cell contents leak
out, and the bacterium dies. These antibiotics do not affect human cells because
human cells do not have cell walls.

Many antibiotics operate by inhibiting the synthesis of various intracellular

bacterial molecules, including DNA, RNA, ribosomes, and proteins. The
synthetic sulfonamides are among the antibiotics that indirectly interfere with
nucleic acid synthesis. Nucleic-acid synthesis can also be stopped by antibiotics
that inhibit the enzymes that assemble these polymers—for example, DNA
polymerase or RNA polymerase. Examples of such antibiotics are actinomycin,
rifamicin, and rifampicin, the last two being particularly valuable in the
treatment of tuberculosis. The quinolone antibiotics inhibit synthesis of an
enzyme responsible for the coiling and uncoiling of the chromosome, a process
necessary for DNA replication and for transcription to messenger RNA. Some
antibacterials affect the assembly of messenger RNA, thus causing its genetic
message to be garbled. When these faulty messages are translated, the protein
products are nonfunctional. There are also other mechanisms: The tetracyclines
compete with incoming transfer-RNA molecules; the aminoglycosides cause the
genetic message to be misread and a defective protein to be produced;
chloramphenicol prevents the linking of amino acids to the growing protein; and
puromycin causes the protein chain to terminate prematurely, releasing an
incomplete protein.

In some species of bacteria the cell wall consists primarily of a thick layer of
peptidoglycan. Other species have a much thinner layer of peptidoglycan and an
outer as well as an inner membrane. When bacteria are subjected to Gram's
stain, these differences in structure affect the differential staining of the bacteria
with a dye called gentian violet. The differences in staining coloration (gram-
positive bacteria appear purple and gram-negative bacteria appear colorless or
reddish, depending on the process used) are the basis of the classification of
bacteria into gram-positive (those with thick peptidoglycan) and gram-negative
(those with thin peptidoglycan and an outer membrane), because the staining
properties correlate with many other bacterial properties. Antibacterials can be
further subdivided into narrow-spectrum and broad-spectrum agents. The
narrow-spectrum penicillins act against many gram-positive bacteria.
Aminoglycosides, also narrow-spectrum, act against many gram-negative as
well as some gram-positive bacteria. The tetracyclines and chloramphenicols
are both broad-spectrum drugs because they are effective against both gram-
positive and gram-negativby antagonizing a wide range of microbes including
bacteria, fungi, and enveloped viruses. The antibiotic activity is thought to be
based on their cationic, amphipathic nature, which enables the peptides to
impair vital membrane functions. Molecular details for such activities have been
elaborated with model membranes; however, there is increasing evidence that
these models may not reflect the complex processes involved in the killing of
microbes. For example, the overall killing activity of the bacterial peptide
antibiotic nisin is composed of independent activities such as the formation of
target-mediated pores, inhibition of cell-wall biosynthesis, formation of
nontargeted pores, and induction of autolysis. We studied the molecular modes
of action of human defense peptides and tried to determine whether they impair
membrane functions primarily and whether additional antibiotic activities may
be found. We compared killing kinetics, solute efflux kinetics, membrane-
depolarization assays, and macromolecular biosynthesis assays and used several
strains of Gram-positive cocci as test strains. We found that membrane
depolarization contributes to rapid killing of a significant fraction of target cells
within a bacterial culture. However, substantial subpopulations appear to survive
the primary effects on the membrane. Depending on individual strains and
species and peptide concentrations, such subpopulations may resume growth or
be killed through additional activities of the peptides. Such activities can include
the activation of cell-wall lytic enzymes, which appears of particular importance
for killing of staphylococcal strains

Mechanism of action

Antibiotics active against bacteria are bacteriostatic or bacteriocidal; that

is, they either inhibit growth of susceptible organisms or destroy them. On the
basis of their mechanism of action, antibiotics are classified as (1) those that
affect bacterial cell-wall biosynthesis, causing loss of viability and often cell
lysis (penicillins and cephalosporins, bacitracin, cycloserine, vancomycin); (2)
those that act directly on the cell membrane, affecting its barrier function and
leading to leakage of intracellular components (polymyxin); (3) those that
interfere with protein biosynthesis (chloramphenicol, tetracyclines,
erythromycin, spectinomycin, streptomycin, gentamycin); (4) those that affect
nucleic acid biosynthesis (rifampicin, novobiocin, quinolones); and (5) those
that block specific steps in intermediary metabolism (sulfonamides,
trimethoprim).

Antibiotics active against fungi are fungistatic or fungicidal. Their mechanisms

of action include (1) interaction with the cell membrane, leading to leakage of
cytoplasmic components (amphotericin, nystatin); (2) interference with the
synthesis of membrane components (ketoconazole, fluconazole); (3)
interference with nucleic acid synthesis (5-fluorocytosine); and (4) interference
with microtubule assembly (griseofulvin). See also Fungistat and fungicide

loped viruses. The antibiotic activity is thought to be based on their cationic,

amphipathic nature, which enables the peptides to impair vital membrane
functions. Molecular details for such activities have been elaborated with model
membranes; however, there is increasing evidence that these models may not
reflect the complex processes involved in the killing of microbes. For example,
the overall killing activity of the bacterial peptide antibiotic nisin is composed of
independent activities such as the formation of target-mediated pores, inhibition
of cell-wall biosynthesis, formation of nontargeted pores, and induction of
autolysis. We studied the molecular modes of action of human defense peptides
and tried to determine whether they impair membrane functions primarily and
whether additional antibiotic activities may be found. We compared killing
kinetics, solute efflux kinetics, membrane-depolarization assays, and
macromolecular biosynthesis assays and used several strains of Gram-positive
cocci as test strains. We found that membrane depolarization contributes to rapid
killing of a significant fraction of target cells within a bacterial culture.
However, substantial subpopulations appear to survive the primary effects on
the membrane. Depending on individual strains and species and peptide
concentrations, such subpopulations may resume growth or be killed through
additional activities of the peptides. Such activities can include the activation of
cell-wall lytic enzymes, which appears of particular importance for killing of
staphylococcal strainsesistance to antimicrobials.

The four main mechanisms by which microorganisms exhibit rwere first

described in France by Ernest Duchesne in 1897. .

1. Drug inactivation or modification: e.g. enzymatic

deactivation of Penicillin G in some penicillin-resistant
bacteria through the production of β-lactamases

Drugs and foreign chemicals that are lipid soluble are converted by enzymatic
processes to compounds of ever-increasing water-solubility until they can be
excreted via one or several of the routes available. Metabolism or
biotransformation and the subsequent excretion of drugs is known as
 “elimination.” Metabolism generally occurs in 2 phases: Phase I induces a
chemical change (most frequently oxidation, but also reduction) hat renders the
drug more conducive to phase II. Phase II is a conjugative or synthetic addition
of a large, polar molecule that renders the drug water soluble and amenable to
renal excretion.
Several aspects of the biotransformation of drugs have direct clinical
significance. These include microsomal enzyme induction and inhibition,
There are several possible consequences of the biochemical transformation of
drugs: 1) inactivation, during which an active drug is converted to inactive
metabolite(s); 2) activation, during which an inactive drug (or pro-drug) is
converted to a pharmacologically active primary metabolite; 3) modification of
activity after the conversion of an active drug to a metabolite that also has
pharmacologic activity; 4) lethal synthesis (or intoxication), in which a drug is
incorporated into a normal cellular metabolic pathway that ultimately leads to
failure of the reaction sequence because of the presence of spurious substrate
(cell death then occurs).
nutritional state, age, disease conditions, and species differences.
Because drug biotransformation is negligible in early life, neonates are much
more sensitive to lipid-soluble drugs than are adults of any species. The
postnatal development of drug-metabolizing enzymes in the liver appears to be
biphasic, consisting of a rapid and nearly linear increase in activity during the
first 3-4 wk, followed by slower development up to the tenth week postpartum;
dose or interval for the very young must be reduced accordingly. Hepatic mass,
hepatic blood flow, and microsomal enzyme activity may decrease in older
animals.
Many disease states impair the normal activity of the hepatic microsomal
enzyme system, which in turn prolongs the half-lives of many drugs. Frank
hepatotoxicity, acute hepatitis, or other extensive liver lesions invariably depress
enzyme activity. Changes in hepatic blood flow, with similar consequences, may
be encountered in congestive heart failure, circulatory shock, and cirrhosis.
Hypothyroidism tends to reduce microsomal enzyme function, and
hyperthyroidism tends to increase activity.
Species variations in the biotransformation patterns of lipid-soluble
drugs are common. Differences in the duration of action of these drugs
in various species frequently can be attributed to differences in their
rates of biotransformation. This must be remembered when either
dosages or withdrawal times are extrapolated from one species to
another.

2. Alteration of target site: e.g. alteration of PBP—the

binding target site of penicillins—in MRSA and other
penicillin-resistant bacteria.

• Enterococcal resistance to vancomycin - different types but

all involve synthesis of altered cell wall precursor side
chain that doesn’t bind vancomycin e.g., D-Ala-D-Lactate
vs. D-Ala-D-Ala
– This mechanism may be plasmid-mediated
• Staphylococcal resistance to semisynthetic penicillins
– Synthesis of a novel penicillin binding protein (2a) with reduced
affinity for methicillin
– Takes over role of other PBPs (which are methicillin susceptible)
3.Metabolism is a step by step modification of the initial molecule to shape it

into another product. The result can be used in one of three ways.

• Stored by the cell.

• Be used immediately, as a metabolic product.
• Initiate another metabolic pathway, called a flux
generating step.

A molecule called a substrate enters a metabolic pathway depending on the

needs of the cell and the availability of the substrate. An increase in
concentration of anabolical and catabolical end products would slow the
metabolic rate for that particular pathway.

Metabolic pathways are composed of a series of biochemical reactions that are

connected by their intermediates: the reactants (or substrates) of one reaction
are the products of the previous one, and so on. Metabolic pathways are usually
considered in one direction (although all reactions are chemically reversible,
conditions in the cell are such that it is thermodynamically more favorable for
flux to be in one of the directions).
 •

Glycolysis was the first metabolic pathway discovered:

1. As glucose enters a cell it is immediately phosphorylated by

ATP to glucose 6-phosphate in the irreversible first step. This is to
prevent the glucose leaving the cell.
2. In times of excess lipid or protein energy sources glycolysis
may run in reverse (gluconeogenesis) in order to produce glucose
6-phosphate for storage as glycogen or starch.

Major metabolic pathways

4. Mechanisms of Drug Action and Resistance

Chemotherapy is the primary means of treating protozoan infections. Successful

chemotherapy depends in a large part on the ability to exploit metabolic
differences between the pathogen and the host. A problem confronting
chemotherapy is the ability of the pathogen to mutate and become drug
resistance. Drugs act by specifically interferring with cellular or biochemical
processes, often called 'targets'. The classic example of a drug target is an
enzyme which is
inhibited by the drug.
Effective drugs will
exhibit a selective
toxicity for the
pathogen as compared
to the host. Rational
drug design seeks to
exploit these various
factors to develop
drugs which are highly
toxic to the pathogen
and at the same time
exhibit minimal toxicity to the host.
 Simplified scheme of folate metabolism. The malaria parasite synthesizes folates de novo,
but cannot utilize preformed folates. Folates participate as co-factors in many biosynthetic
processes. Of particular note is the synthesis of thymidylate (dTMP) which is needed for
DNA synthesis. The two primary targets of antimalarial drugs which target folate metabolism
are denoted with the boxed arrows.

Applications
Antibiotic resistance is an important tool for genetic engineering. By
constructing a plasmid which contains an antibiotic resistance gene as well as
the gene being engineered or expressed, a researcher can ensure that when
bacteria replicate, only the copies which carry along the plasmid survive. This
ensures that the gene being manipulated passes along when the bacteria
replicates.
The most commonly used antibiotics in genetic engineering are generally
"older" antibiotics which have largely fallen out of use in clinical practice.
These include:

• ampicillin
• kanamycin
• tetracycline and chioramphenic.

Refrences

www.google.com
www.yahoo.com
www.knowledgestorm.com
www.nytimes.com
11th class comprehensive
12th class biotextbook

Rosen P, Barkin RM, eds. Emergency Medicine:

Concepts and Clinical Practice.
Tintinalli JE, Krome RL, Ruiz E. Emergency
Medicine: A Comprehensive Study Guide. 4th ed.