Biology

1) Ans.) Serologically cross-reactive matrix protein and nucleoprotein, are used in differentiating type-A Influenza virus from the internal proteins of type-B and type-C strains.

2) Ans.) Nucleotide sequencing of the 5-prime and 3-prime ends of the genomic RNA of strains of types A, B, and C revealed a high degree of conservation of the terminal 10 to 20 nucleotides which act as the common progenitor for all the three viral subtypes.

3) Ans.) 1.Infrequent association of virus-C with disease in man, frequent association of virus-B with the disease in man was observed and virus A is associated with pandemic influenza in man.
2. A virus has wide host range (man, pigs, horses and birds).
3. Un-like influenza-A viruses, type-B and type-C do not appear to undergo rapid major genetic or antigenic changes. Virus-C does not have neuraminidase gene.

4) Ans.) Recombination of the genes that lead to the synthesis of surface proteins (which play important role in giving antigenic specificity to the virus in causing disease) occurring between different strains of subtypes is called re-assortment. If this recombination occurs significantly and suddenly it is called as antigenic shift. But if recombination of genes leading to non-surface proteins occurs non-significantly within the viral subtypes it is called as antigenic drift.
Re-assortment of genes Recombination of genes between different strains of same subtype or between different subtypes either leading to synthesis of surface proteins or non-surface proteins is called re-assortment of genes.

Antigenic shift Dramatic occurrence of recombination of genes that code for surface antigen proteins among different strains of the viral subtypes.

5) Ans.) If the serum samples of people suffered from a recent pandemic caused by a strain has cross-reacted with another strain that has appeared quiet a few years back, it is an indirect evidence to say that there are a limited number of viral antigens (hemagglutinin genes) that have potential to cause epidemics and pandemic, which keep recycling in the strains although the years. This is called antigen recycling.

6) Ans.) Three basic features appear to be conserved in type-A Influenza virus hemagglutinin which shows great degree of variability. The regions of amino-acid sequence that are conserved are amino-terminal end of Hemagglutinin2 subunit, carboxyl terminal hydrophobic tail of Hemagglutinin2 subunit and many cysteine residues along the entire sequence. These three regions are found to be conserved which means not variable or retained in the evolutionary process. These conserved sequences are responsible to carryout series of common important biological functions and penetration of virus into cells.

7) Ans.) Three HemagglutininA subtype viruses H1, H2 and H3 can infect humans and two NeuraminidaseA subtype viruses N1 and N2 can infect humans.

8) Ans.) Yes, The nucleotide substitutions that were studied by sequence data in the hemagglutinin genes of different H3 subtypes were found to be roughly linear and cumulative. If the nucleotide substitutions occurred are conserved although the evolution of the strains one after the other it is called as linear. If the substitutions continue to increase and get conserved then it is called cumulative.

9) Ans.) 1. Immune response in some patients that led to favor the selection of variants through several cycles of virus replication.
2. The sheer number of people infected with influenza viruses during an epidemic may result in the formation of large numbers of variants.
3. The change of one or more amino acids in the influenza virus hemagglutinin may change the antigenic character of the virus sufficiently, whereas a similar number of amino acid changes may not provide such a selective advantage for other kinds of viruses.

10) Ans.) With the help of Recombinant DNA techniques effective vaccines are generated. Stable attenuated (half killed) virus that can be used as live virus vaccine can be constructed. Influenza virus strains may be constructed which contain altered antigenic determinants (antigenic sites). Viral antigens or portions of viral antigens in bacterial or yeast cells for use as killed virus vaccines can be produced by genetic engineering methods. Synthesis of modified antigens that exhibit determinants cross-reactive among strains (of various subtypes) may lead to preparation of vaccines that represent a significant improvement over conventional vaccines. Effective influenza virus vaccine takes advantage of the chemical synthesis of specific peptides. These peptides have been shown to be immunogenic and synthesis of the right amino acid stretch derived from known viral sequences or construction of a suitable cross-reactive immunogen containing multiple determinants may lead to the ultimate influenza virus vaccine.

Table2
This table describes about the total length of nucleotides in hemagglutinin gene, amino-acid length in hemagglutinin1 subunit and hemagglutinin2 subunit of H1 (1934), H2 (1957) and H3 (1968) viral Influenza-A subtypes. So the nucleotide lengths were 1778, 1773 and 1765 in H1, H2 and H3 viral Influenza-A subtypes respectively. In the same way, amino-acid lengths of hemagglutinin1 subunit in all the three subtypes were 326, 324 and 328 respectively. Again the amino-acid lengths of hemagglutinin2 subunits in the three subtypes were 222, 222 and 221 respectively.

Table3
Percentage nucleotide conservation in hemagglutinin1 and hemagglutinin2 subunits in
H1 and H2 viral Influenza-A subtypes           61         72
H1 and H3 viral Influenza-A subtypes           45         58
H2 and H3 viral Influenza-A subtypes           45         57
Percentage amino-acid conservation in hemagglutinin1 and hemagglutinin2 subunits in
H1 and H2 viral Influenza-A subtypes            58         79
H1 and H3 viral Influenza-A subtypes            35          53
H2 and H3 viral Influenza-A subtypes             36           50
Nucleotide sequence of the haemagglutinin gene of a human influenza virus H1 subtype

Abstract
1.H1 subtype of human influenza virus found to be dominant in this century was found to be co-circulating with variants of H3 subtype.
2.The sequencing of haemagglutinin gene of an early H1 subtype (strain APR834) was done by recombinant DNA methods and di-deoxy methods.
3.The antigenic site was defined at amino-acid residue 160.
4.After comparison of amino-acid residues among the different subtypes H1 and H2 were found to homologous to one another than all others.

-Influenza virus consists of 8 RNA genes ( 10 proteins, in which band 4 is meant for haemagglutinin gene.
-Methodology of sequencing 1. Generate restriction fragments of double-stranded DNA from RNA of gene4 by reverse transcriptase. 2. These fragments were cloned into the bacteriophage M13 vector and sequenced by sangers method. 3. Then all the sequences of isolated clones were overlapped to deduce unique sequence for the haemagglutinin gene. 4. An indirect RNA sequencing method was also used.
-Haemagglutinin gene, in its mRNA sense coded into amino-acid sequence consisted of 17-residue hydrophobic signal peptide, a haemagglutinin1 subunit (326 residues) and a haemagglutinin2 subunit (222 residues) separated by Arginine residue.
-Haemagglutinin2 subunit contains 14-residue N-terminal hydrophobic sequence and hydrophobic sequence.
-Disulphide bridges or bonds existed within the subunits but at C14 and C466 they are present between the subunits.
-A common antigenic variant of influenza-A, APR834 was identified to have changed serine to leucine at residue position 160 which is C to U transition at nucleotide position 553.
- The antigenic variance at position 160 of H1 subtype virus was homologous to 157 position of H3 subtype.
Table1

Percent amino-acid conservation in haemagglutinin1 haemagglutinin2 subunits in
H2 and H1 viral sub-types         58             79
H3 and H1                35            53
H3 and H2                36            50   
H7 and H1                33             51
H7 and H2                35              65

Percent nucleotide conservation in haemagglutinin1 haemagglutinin2 subunits in
H1 and H2 viral subtypes        61              72
H3 and H1                45              58
H7 and H1                44              58
H3 and H2                45               57
H7 and H2                46               59
H7 and H3                45               66

-Amino-acid conservation was more between H1 and H2 viral subtypes than between all other subtypes.
-Homology of all haemagglutinin subtypes indicates that they might have evolved from common ancestor.
- From the extent of antigenic drift of haemagglutinin1 subunit observed in H3 series ANT6068,
AMemphis10272 and AVictoria275, it was estimated that 1 amino-acid change per year occurred.
- So by applying this drift, 42 difference in amino-acid sequence of haemagglutinin1 subunit of H1 and H2 subtypes might have been derived from the assumption that H2 subtype has evolved from H1 subtype in a 42-year period.
- Drift of H2 subtype from H1 subtype (1933) took place at 1957 and no other strains were found to be existing between these two affecting man. Even in avians and pigs also no intermediate strains are found in the last 48years.
- H1 and H2 viral subtypes were identified to be more evolutionarily related than to H3 and H7 subtypes though the evolutionary distance was not clearly estimated.

Future
It would be more informative further for everyone in this regard to sequence completely some of eight other known mammalian or avian haemagglutinin molecules because subtypes that are distantly related which may have been undetected by traditional methods used to classify haemagglutinin subtypes can also be checked for homology.

1.Bordettella pertussis gram negative coccobacilli which causes whooping cough
Symptoms of the disease Prolonged dry cough
Infective source inhalation of bacterium in droplets released from an infectious person
Diagnosis fluorescent antibody staining of smears from nasopharyngeal swabs and serological tests
Treatment administration of penicillin, tetracycline or chloramphenicol
Prevention Vaccination with DPT vaccine

2. Mumps virus infectious disease caused by Rubula virus
Symptoms of the disease swelling of the salivary glands, especially the parotids, and sometimes of the pancreas, ovaries, or testes
Infective source spread by direct contact, airborne droplet nuclei
Diagnosis Culture, Serology tests of IgG and IgM.
Treatment Orchitis Symptomatic relief, Prednisone 60 mg qd tapered over 7-10 days
Prevention MMR Vaccine after age 1 year

3.Rubeola virus an RNA virus of the genus Morbillivirus, family Paramyxoviridae, that causes measles in humans
Symptoms of the disease cold like symptoms, running nose, sneezing, red eyes
Infective source spread by direct contact or by airborne respiratory drop-lets.
Diagnosis distinctive clinical features and serological tests
Treatment none
Prevention vaccination

4.Rubella virus Rubella, commonly called German Measles caused by the Rubella virus
Symptoms of the disease headache, malaise, anorexia, low-grade fever.
Infective source transmitted through contact with the blood or nasopharyngeal secretions.
Diagnosis Cell cultures of the throat, blood, urine, and cerebrospinal fluid can confirm the virus presence.
Treatment Treatment consists of aspirin for fever and joint pain.
Prevention Immunization with live virus vaccine RA273.

5.Common cold virus minor contagious infection caused by a number of different viruses (e.g. rhinoviruses, coronaviruses).
Symptoms of the disease cold, low grade fever, body pains, running nose
Infective source droplets of the infected people, sneezing
Diagnosis distinctive clinical features
Treatment antiviral agents. Not specific
Prevention avoid contact with the infected people

6. Haemophilus influenza  gram negative rod shaped, encapsulated of type b
Disease name bacterial meningitis. It causes diseases in many organ systems but usually attacks the respiratory system.
 Symptoms of the disease influenzae infects the larynx, trachea, or bronchial tree, it leads to irritable cough, dyspnea, mucosal edema, and thick, purulent exudate.
Infective source Transmission occurs by direct contact with secretions or by respiratory droplets.
Diagnosis bacterial culture or latex particle agglutination.
Treatment cefotaxime and ceftriaxone are the elected antibiotics
Prevention Vaccination   

7. Haemophilus  aegyptius  conjunctivitis causing gram negative rod shaped bacteria
Symptoms of the disease red eyes, watering of eye, low grade fever
Infective source air-borne infection
Diagnosis culture of bacteria from droplets
Treatment none
Prevention none

8.Coxiella Burnetii Q-fever causing gram negative bacterium
Symptoms of the disease fever, myalgia , headache,
Infective source inhalation of dust contaminated with bacterium from animal faeces, urine or milk
Diagnosis serological test
Treatment administration of chloramphenicol and tetracycline
Prevention vaccination and pasteurization of sheep and cow milk

9.Strepococcus pyogenes A species of gram-positive, coccoid bacteria isolated from upper respiratory tract of humans causes scarlet fever
Symptoms of the disease sore throat, swollen lymph glands, fever and headache
Infective source spread through contaminated droplets from an infected person
Diagnosis marked leukocytosis with neutrophilia and increased ESR in blood test
Treatment administration of penicillin or erythromycin
Prevention Individuals infected should be isolated and treated

10.Pneumocystis carinii (jiroveci) Pneumocytosis is caused by yeast like fungus
Symptoms of the disease fever, non-productive cough, weight loss and night sweats
Infective source respirtatory droplets
Diagnosis chest x-ray, lower arterial oxygen level,histological identification in sputum
Treatment trimethoprim,sulfmethoxazole are used
Prevention avoid close contact with infected people

11.Klebsiella pneumonia Gram-negative, capsulated, gas-producing rods associated with urinary and respiratory infections in humans.
Symptoms of the disease bacteremia, pneumonia, urinary tract infections
Infective source found in intestinal tract and it is opportunistic pathogen
Diagnosis chest x-ray, culture specimens of sputum and blood
Treatment resistant to multiple antibiotics. Treatment depends on the organ system involved.
Prevention vaccination

12.Variola virus smallpox is an acute, highly contagious infectious disease caused by the poxvirus variola.
Symptoms of the disease chills. High fever, headache, sore throat, lesions  on mucous membranes of mouth, throat and respiratory tract.patient soon develops skin lesions progressing from macular stage to pustular stage
Infective source respiratory droplets or dried scales of virus-containing lesions.
Diagnosis complement fixation to detect virus or antibodies in blood, culture of variola virus.
Treatment  antimicrobial therapy, symptomatic treatment of lesions with antipruritics, starting during the pustular stage.
Prevention Vaccination

13.Herpes virus varicellae ds- DNA virus which belongs to Herpes simplex virus Symptoms of the disease pruritic rash, slight fever, anorexia,
Infective source direct contact with respiratory secretions
Diagnosis based on clinical signs and serological tests
Treatment anti-itch creams, acyclovir, zovirax
Prevention immunization with varicella virus vaccine

14.Streptococcus pneumoniae gram positive cocci occurring in pairs or chains.
Symptoms of the disease pneumonia, sinusitis
Infective source respiratory droplets
Diagnosis sputum gram staining
Treatment antimicrobial therapy varies analgesic is given to relieve pleuritic chest pain
Prevention vaccination

15. Neisseria meningitides gram negative diplococci causes meningococcal meningitis
Symptoms of the disease nausea, vomiting, headache, fever, stiff neck, sleepiness
Infective source transmission through close contacts
Diagnosis lumbar puncture and serological test
Treatment ceftriaxone is generally prescribed. steroid medication is also used
Prevention immunization with meningococcal C conjugate vaccine,

16.Corynebacterium diphtheria diphtheria causing gram positive pleomorphic bacteria
Symptoms of the disease soar throat, fever, neck swelling, difficulty in swallowing
Infective source Transmission occurs person-to-person spread from the respiratory tract.
Diagnosis serological test and culture of the bacterium on specific medium
Treatment penicillin and erythromycin are generally prescribed
Prevention immunization with diptheria toxoid containing vaccine

17.Mycobacterium tuberculosis bacilli phylogenetically positive but stains acid fast which causes tuberculosis.
Symptoms of the disease Persistent cough, chest pain, weight loss, chills, fever
Infective source droplet nuclei produced when infected persons cough or sneeze.
Diagnosis chest X-ray, sputum test, TB skin test (Mantoux test) or purified protein derivative (PPD) test.
Treatment combination of drugs isoniazid (INH) and rifampin-6months, and pyrazinamide and ethambutol-first two months of treatment.
Prevention identifying infected individuals early  especially those who run the highest risk of developing active disease and treating them

18. Mycobacterium leprae gram-Positive Asporogenous Rods, Regular, causes. leprosy (Hansens disease)
Symptoms of the disease symmetric skin lesions, nodules, plaques, thickened dermis,
Infective source  respiratory droplets
Diagnosis Biopsies of skin lesions, Peripheral nerve biopsy or smears of the skin or of ulcerated mucous membranes Blood tests show increased erythrocyte sedimentation rate decreased albumin, calcium, and cholesterol levels.
Treatment Dapsone, Dapsoderm-X, clofazime and lamprene
Prevention  Avoid close contacts with patients with untreated.

19.Legionella pneumophila aerobic gram-negative rod survives well in protozoa causes legionellosis
Symptoms of the disease high fever, non-productive cough, neurological manifestations, bronchopneumonia
Infective source air-borne spread from environmental reservoir
Diagnosis rapid test kit using urine to detect antigens
Treatment supportive measures and administration of erythromycin or rifampin
Prevention identification and elimination of environmental protozoans

20.Mycoplasma pneumoniae gram negative pleomorphic bacteria
 Symptoms of the disease tracheobronchitis, pneumonia, fever, non-productive cough
Diagnosis chest X-ray showing infiltrates, sputum smear demonstrating acute inflammatory cells
Infective source spread involves close contact and sometimes air droplets
Treatment tetracycline or erythromycin
Prevention None

21.Toxoplasma gondii toxoplasmosis is caused by the protozoan
Symptoms of the disease swollen lymph nodes, muscle aches and pain
Infective source partly cooked meat, ingestion of contaminated cat faeces
Diagnosis polymerase chain reaction technique
Treatment clindamycin and spiramycin
Prevention avoid the contaminated drinking water and partially cooked meat

22. Aspergillus species These saprophytic molds cause aspergillosis
Symptoms of the disease
Infective source air-borne spread of conidiospores
Diagnosis chest CT, new rapid stain techniques and serum ELISA
Treatment voriconazole, amphotericin B cholesteryl sulfate complex
Prevention none.

23.Histoplasmosis capsulatum it is a fungus which causes Histoplasmosis
Symptoms of the disease respiratory symptoms, malaise and fever
Infective source  contact with bird or bat droppings
Diagnosis chest x-ray
Treatment nizoral and ketoconazole
Prevention Avoid areas that may harbor the fungus.

24.Cytomegalovirus (CMV) ds-DNA virus belongs to genus of the family herpesviridae
Symptoms of the disease  fever, swollen lymph glands, retinitis, pneumonitis
Diagnosis CMV antibody blood test, viral culture, quantitative antigenemia assay
Infective source The virus, a member of the herpesvirus family, is found in saliva, urine, and other bodily fluids. Because it is often found in semen as well as in cervical secretions, the virus can be spread by sexual contact it also can be easily spread by other forms of physical contact such as kissing.
Treatment None
Prevention Drainage and secretion, and pregnant women precautions, should be instituted for hospitalized patients known to be shedding CMV.

25. Erythema infectiosum (fifth disease) Parvovirus B19 with single stranded DNA
 Symptoms of the disease facial rash, body rash, aplastic crisis, chronic marrow suppression
Diagnosis Hematocrit and reticulocyte count in patients with aplastic crisis, Parvovirus DNA by PCR testing
Infective source  respiratory droplets or blood, transplacental.
Treatment symptomatic treatment, pain relief medications.
Prevention Pregnant women should avoid contact with Parvovirus

26.RSV (Respiratory Syncytial virus) negative ss-RNA virus belongs to family paramyxoviridae cause rhinitis and laryngitis
Symptoms of the disease fever, cough, rhinitis and nasal congestion
Infective source hand contact and respiratory secretions of humans
Diagnosis directigen RSV or Test-Pack RSV rapid test kits.
Treatment inhaled ribavirin
Prevention isolation of RSV- infected individuals

27.Coccidiodes immitis Coccidioidomycosis is caused by this fungus
Symptoms of the disease rash, myalgia, malaise, flu like symptoms
Infective source generally found in soil and transmission occurs through skin
 Diagnosis VDRL test, blood test, urinalysis, chest x-ray
Treatment amphotericin B, fluconazole
Prevention none

28.SARS (coronavirus) Severe acute respiratory syndrome (SARS) caused by virus which belongs to family coronavirus
Symptoms of the disease chills, fever, shivering, body aches, muscle aches
Infective source droplet transmission from infected people.
Diagnosis sputum gram stain and culture, chest x-ray, WBC count
Treatment oral steroids, ribavarin, oseltamivir
Prevention avoid droplet exposure and use surgical masks.

29.Hantavirus ss-RNA negative strand virus belongs to genus of the family Bunyaviridae causing Hantavirus Pulmonary Syndrome.
Symptoms of the Disease Progressive Tachypnea, Cough productive of copious secretions
Infective source Inhalation of aerosolized rodent feces or Saliva, No identified cases of person to person transmission
Treatment Drug therapy includes vasopressors, such as dopamine or epinephrine, for hypotension. Ribavirin in aerosol form has been used for children
Diagnosis Sin Nombre Virus serologic titers
Prevention Eliminate rodent populations in and around buildings. avoid rodent exposure

30. Ebolamarbug virus A genus in the family Filoviridae consisting of several distinct species of Ebolavirus. These viruses cause outbreaks of a contagious, hemorrhagic disease (hemorrhagic fever) in humans
Infective source Infected blood, Body fluid, Body tissue
Symptoms of the disease fever, chills, myalgia, malaise, severe abdominal pain.
Treatment None
Diagnosis serological test demonstrate neutrophil leukocytosis, hypofibrinogenemia, thrombocytopenia, and microangiopathic hemolytic anemia.
Prevention Precautions to avoid contact with patients body fluidssecretions.

A. summary of structurefunction and how the cardiovascular and respiratory systems co-work.   Introduction
   
Proper functioning of the cardiovascular system is very dependent on the respiratory system. Equally, the functions of the respiratory system cannot be complete without the cardiovascular system.
Thesis statement the cardiovascular system and the respiratory system co-work to accomplish vital and complex roles in the body thus making it very hard to make a robot that would duplicate their roles.

Cardiovascular system   
The cardiovascular (circulatory) system is made of three main components the blood, blood vessels and the heart. The blood is made of several components including erythrocytes which contain hemoglobin thus helping in oxygen transport. The lymphocytes on the other hand are responsible for providing the body with immunity by T cells specifically killing infected cells directly whereas the B lymphocytes produce antibodies. Thrombocytes (platelets) are responsible for blood clotting whereas leukocytes phagocytose invading microorganisms. The plasma is the fluid component of the blood primarily made of water with only a small percentage containing dissolved nutrients, wastes, enzymes and hormones. The blood plasma carries the function of keeping a constant body PH in addition to helping in blood clotting through fibrinogen component. Blood plasma is also important in maintaining body immunity through the globulin component in addition to transporting molecules such as cholesterol (PATTS, 2000).
   
There are three types of blood vessels veins, arteries and capillaries. The vascular system is distributed through out the body with the arteries and veins arranged parallel to each other whereas the capillaries form a web-like connection. The arteries are highly elastic and are generally strong. Blood is transported at high pressure in the arteries as it leaves the heart to other parts of the body. The strength and elasticity of these vessels allows for this. As they move further from the heart, the arteries split into thinner vessels progressively into arterioles. The largest artery is the aorta with branches such as the coronary arteries (right and left) as well as descending and ascending aorta (Cardiology Channel, 2008).
   
As the arterioles branch further, they form the web of capillaries which are microscopic in size. The capillaries have a thin wall that allows substances to cross into the surrounding tissues.  The capillaries then connect to venules whose later connection forms the veins. Veins are not as strong as the arteries as their inner walls are thinner than those of the arteries and blood moves through them at a low pressure. The veins therefore contain valves to prevent blood from flowing back. Connecting the veins directly to the heart is the inferior and superior vena cava. 
   
It is important to note that arteries carry blood away from the heart at a high pressure and the blood is rich in oxygen. However an exception occurs with pulmonary artery that takes blood to the lung for oxygenation. The veins on the other hand transport deoxygenated blood back to the heart save for pulmonary vein that carries blood oxygen rich blood from the lungs to the heart (Cardiology Channel, 2008).
   
The heart is a major organ in the cardiovascular system and therefore it has a complex structure and function. Close to the size of a mans fist, the heart is flanked by the lungs with its greater part (two-thirds) reclining to the left of the thoracic cavity midline. The heart is internally made up of four chambers with the atria forming the upper chambers and the ventricles forming the lower chambers. The major division of the heart is the right and the left side of the heart. Blood coming from all parts of the body through the inferior and superior vena cavae enter the heart via the right atrium and into the right ventricle through the tricuspid valve. From the right ventricle, it s pumped into the lungs through the pulmonary artery. After oxygenation, blood is transported into the left atrium through the pulmonary vein passing through the mitral valve. The muscular left ventricle then pumps blood at high pressure into all parts of the body through the aorta.  
   
The circulatory circuit is made of the pulmonary circulation involving blood circulation around the heart and the lung and the systemic circulation arising from the heart to all parts of the body. In the systemic circulation, the aorta is the major artery. It is subdivides into aortic arch that supplies the head, the ascending artery supplying the abdomen and the descending aorta supplying the body extremities. The heart itself is supplied by the coronary arteries (PATTS, 2000).

Respiratory system   
The respiratory system is primarily responsible for facilitating gaseous exchange and eventually maintaining a stable body PH. located in the thorax, it is subdivided into the upper and the lower respiratory system. The upper respiratory system is made of the nose, the nasal cavity, mouth cavity, sinuses, larynx and trachea. These are the upper air passages with the mouth and the nasal cavity conditioning air appropriately which may include moisturizing air and warming air among other functions. Control of moisture and temperature in the nasal cavity is facilitated by the mucous membranes thereof. The lining of the airways not only contains mucus but also has cilia that filter air of any particles. At the pharynx, the epiglottis ensures that food does not pass into the trachea through the larynx. Air passes the larynx into the bronchi (right and left bronchi connecting to each lung). Important to note is that the bronchi never collapse due to reinforcement by rings of cartilage. The bronchi subdivides further into narrow tubes called the bronchioles which connect to the alveoli of the lungs. The alveoli are small sacs which are clustered to form grape like shape. Surrounding the alveoli is an extensive network of capillaries that facilitates gaseous exchange. 
  
The lungs are located in the thoracic cavity and they are a pair of lobular organs divided into an upper lobe, a lower lobe (the left lung) and a middle lobe (the right lung). Surrounding the lungs is the ribcage made of ribs and intercostals muscles whose movements facilitate exhalation and inhalation. Distal to the lung is the diaphragm (a strong muscle) which moves up and down also thus facilitating ventilation. During ventilation, air passes through the conduits into the alveoli in the lungs where exchange of gases occurs. 
The cardiovascular and respiratory systems
   
The lungs form the meeting point of the cardiovascular (specifically the blood) and the respiratory system. The function of the blood is to supply oxygen to all cells of the body whereas the respiratory system oxygenates blood by removing carbon dioxide from the blood. This happens in the alveoli which are surrounded by plenty of capillaries. Deoxygenated blood that has been brought to the lungs from the heart through the pulmonary artery reaches the capillaries in the alveoli. Since the concentration of carbon dioxide is higher in the blood than in the alveoli, it diffuses into the alveoli and removed outside the body through the air passages. On the other hand, air in the alveoli is highly concentrated with oxygen than in the capillaries and hence oxygen diffuses into the capillaries and eventually blood is oxygenated. It is then transported through the respective cardiovascular pathways into every part of the body (Wissman, 2007).
   
As earlier mentioned, blood plasma maintains a constant PH. This is facilitated by the respiratory system whereby removal of carbon dioxide increases alkalinity of blood. The physiological PH should be about PH 7.4. Carbon dioxide in the blood exists in form of a weak carbonic acid which tends to lower blood PH. Removing the carbon dioxide through the respiratory system therefore raises PH to the homeostatic level (Pulmonology Channel, 2009). Interestingly, the cardiovascular system also supplies the respiratory system and therefore they are not independent.

Can a robot replace the cardiovascular and respiratory systems   
The field of science and technology has no impossibilities. Never would one have thought that there would be systems that would take up some vital roles only believed to be confined to the human species.  Technology has led to the existence of robots that perfectly take up the role of the muscular and skeletal systems. There are robots with the ability to open not only doors but wheel chairs. Others have mimicked the nervous system as robots can sense variations in pressure. Others have been made to perform intelligent tasks such as surgery. In spite of all these advancements, there has not been made a robot that would duplicate the role of the cardiovascular and respiratory system (NewScientist, 2010).
   
The functioning of these two systems is quite complex and is not mere mechanical roles. The complex physiology involved in exchange of gases as well as the role of the blood cells may be quite hard to mimic. There is however no doubt that there are machines mimicking organs in these systems. Artificial hearts and lungs have been made and it is also possible to have systems that would purify blood and ensure homeostasis as enabled by these systems. Building on these and other advances in technology, there is high possibility that soon there will be robots duplicating the cardiovascular and respiratory systems.
   
The cardiovascular system is vital in ensuring blood circulation and the subsequent supply of oxygen and removal of waste products from the cells. The blood is also pertinent in defending the body from infections as facilitated by the blood cells. On the other hand, the respiratory system comprised of the upper and lower respiratory system is a vital system in ventilation and co-works with the cardiovascular system to ensure blood oxygenation and overall maintenance of homeostasis. Though no robot has been made to take up these roles, there is no doubt that the future can bring this to a reality.

The bacterial structures that enable these bacteria to be packaged include that both bacteria produce spores and have protein crystals in addition to the cell wall. Moreover, these bacteria are monomorphic which means they maintain a sole shape and hence they can be identified and packaged with ease. Bacillus thuringensis (Dipel) is used in controlling and killing of larvae of insects like mosquitoes, beetle, black flies, moth and many more. When the insect eats the toxic crystals from the pesticide, the toxin gets activated since their digestive system has got alkaline PH. Consequently, the toxin inserts within the cell membrane of the gut of the insect developing a pore and then swelling, followed by lysis and lastly killing the insect (Madigan, 2005). B. subtilis (Kodiak) is used in controlling of several leaf spot and after harvest diseases because it has ability of forming endospores and hence aiding long storage and comparatively simple commercialization of seed products. For instance, it is used in protection of cotton and some other crops from seedlings diseases (Madigan, 2005).

B. Transport of glucose molecules across the plasma membrane  
Glucose molecule is gotten from a starch polymer through facilitated diffusion whereby the molecules move across the cell membrane through particular transport proteins that are rooted in the cellular membrane. The cell transports the glucose in that the glucose binds to a glucose transporter protein to the plasma membrane of a given cell. As a result, the transporter protein changes its conformation and the release of glucose follows. The glucose inside the cell is instantly phosphorylated (Fitzgerald, 2006). After this, the transporting protein binds to the receptor on the plasma membrane and this conveys a signal to raise the quantity of glucose transporter protein on the cells surface. This then raises the quantity of glucose conveyed from the outside of the cell to into the cell (Fitzgerald, 2006).

A Complex Disease - Malaria can be said to be the most prevalent and the most pernicious. Caused by a Protozoan parasite, Plasmodium species (four species), the disease causes fatality in nearly 2.7 to 3 million human individuals most of whom are children below 5 years (Spelman, 2009 p.1). Simply the disease causes high fever due to medical immunity related disturbances in the body. The only reliable method available for confirmed diagnosis of this disease is microscopic observations of parasite cells in the blood. The parasite gets into the blood due to bite of a mosquito vector, Anopheles sps. Thus the disease is complex because it lives both in mosquitoes and men showing many intermediary distinct stages in its life cycle.
   
Control of this disease has become more demanding because the previous successfully used methods for control have become unsuccessful because both the parasite and the vectors have developed resistances to their respective controlling chemicals such as chloroquinine and DDT. The infected mosquito vector populations of Plasmodium can neither be disinfected nor killed because of their all-pervasive nature and universatility. The disease is asymptomatic. Even the existing diagnostic method can give misleading andor erroneous negativespositives.

Inadequate Treatment  Chloroquinine was the historical and safe method for treatment but Plasmodium has developed resistance to this drug. Post-quinine based drugs, Artemesinin, an alkaloid from Artemisia spp., singly initially and then in combination with other drugs were the used. But unfortunately the parasite has acquired immunity to even these.
   
Since then aggressive researches rapidly adopting the latest molecular biological technologies on different aspects of the pathogenesis biology of the parasite both in the vector and the host have been making just optimistic claims about the potentials of the drugs still to be developed on the controls. The different parasite stages in liver and blood of the host, and mouth-saliva and gut of mosquitoes only confound the directions of treatment complexity. Recent interest in the development of blocking of transmission of the parasite from hosts to vectors is only clearing the known and unknown gaps in the knowledge. Transgenic approaches have been actively investigated in spite of many environmental, ethical and regulatory hurdles. Yet the optimism remains                          

Just by developing science and technologies for transmission prevention between a vector to host and vice versa is unlikely to contribute to sustained control of malarial disease. Based on the confidence in the past success of quinine-based and artimesinin (a sesquiterpene lactone) - based, both of which are plant derived molecules, there has been a silent but steady interest in exploration of other potential plants as sources of other molecules from the rich plant bio-diversity. Over 1,277 plant species from 160 families listed that have been used to treat malaria (Willcox 2004, cited by Spelman, 2009).  For exploring anti-plasmodium herbal parts and extracts procedures used may be as follows. 

Plant Parts and Extracts  Different promising parts of different plants would be used on the basis of existing use literature in different folk and ethic medicine (Bero et al., 2009 p 1). Based on their review of literature it becomes clear that plant species of Asteraceae, Caesalipiniaceae and Leguminoasae contain a wide variety of bio-molecules that have been reported to be showing high (IC50 d 2 mM) in vitro activity against various strains of Plasmodium falciparum ((Bero et al., p. 1428). They have highlighted different potential compounds for development towards antiplasmodial drugs. This paper should give good lead in this context. Plants and their parts would also be based on availability of potential anti-plasmodial molecules available. In the traditional medicine systems most of the time water extracts. But in this significant paper there are very valuable antiplasmodial compounds which are lipophilic and therefore insoluble aqueous solutions. 
   
Recently the need concerted efforts in such directions have provided workable techniques for bioassay-based fractionation and separation of the crude extracts. For instance,   Cimanga et al (2006) reported that among ethanol, dichloroethane and petroleum ether the latter solvent yielded potent molecules for above 60 reduction in parasitemia in vivo rodent tests to Plasmodium berghei.
   
Urgency of a safe cheap and potent antiplasmodial molecule demands a rapid development of an alternative antimalarial drug. Recently a technique called the high throughput screening has been developed for testing accurately effective molecules in short time in relation to many other major and serious human diseases.

Research Proposal  This research proposal on screening potential antiplasmodial molecules from the richly documented plant sources ( Bilia et al, 2005 Canoto et al., 2003 Kaur., 2009 Mambu et al., 2008 Saxena et al., 2003 and  Schwikkard and van Heerden, 2002). 
   
Efforts will also be made on the mechanism of action of some most effective molecules on the malarial parasite based on current techniques and literature.

Objectives 
To screen different plant species known in the folklore and ethnic medicine for malaria treatment.
Some selected solvents known for extraction of specific anti-malarial molecules (example different kinds of alkaloids) would be used for extracting different parts sequentially and sequentially. Standardize the best and easiest combinations for bio-assay tests on in vitro and in vivo anti-plasmodial activity

The effective solvent extracted antiplasmodial molecules would be tested on a wide range of mainly chloroquine resistant Plasmodium falciparum strains as well as those of chloroquinine sensitive categories.
In many instances such promising effective molecules would also be tested for other major animal and human diseases such as cancers and HIV, Flu and so on because some of them have shown good results.

Approaches  From the literature survey it becomes clear that the major effort has been to rely on various popular folk and ethnic medicinal plants used in different geographic boundaries. Incidentally it is mostly in those areas that malarial disease is most prevalent and serious. Several efforts on identification of their respective potential anti-plasmodial molecules and their effective anti-plasmodial concentrations have been done on single molecules in nearly all the reports.

In this work the efforts will be towards combining these molecules at their optimum tested concentrations in different permutations and combinations for faster better efficacy at cheap costs without having to resort to further chemical modifications for better efficacies.

Haemolysis which refers to the rupture of erythrocytes and the discharge of the respiratory pigment, hemoglobin and other intracellular contents into the plasma is a very significant process in the clinical pathology of blood. This process which can occur either in vivo or in vitro can have detrimental effects on the health of an individual. In vivo haemolysis which occurs inside the body can cause anemia while in vitro haemolysis which occurs outside the body can have significant but unwanted effects particularly on medical tests as it can result to inaccurate results because of the inclusion of the contents from haemolysis of red blood cells in the plasma. The process of heamolysis is sometimes attributed to the features of the red blood cells in relation to cell osmosis and permeability. It is argued that one of the reasons as to why red blood cells rupture is because of the extracellular medium (plasma) being more dilute than the intracellular cells of the red blood cells. This causes the red blood cells which are higher in concentration to draw water from the surrounding plasma causing them to swell. If they are not strong enough they could burst liberating hemoglobin into the plasma. Based on this concept, red blood cells have been identified by researchers as having several features that are convenient for them to be used in studying the osmotic and permeability relationships of cells.

Osmotic pressure, which refers to that pressure which must be exerted on a solution to prevent water from the surrounding medium from entering it through a semipermeable membrane, is very important in preventing cells from bursting and resulting to processes such as haemolysis in the case of red blood cells. This pressure is often referred to as hydrostatic pressure which means water stopping pressure.

Osmotic and permeability relations of cells depend on the concentration of the blood plasma which is the extracellular solution in which cells exist. When the plasma is isotonic (same concentration as inside the cells), the cells will neither lose nor draw water implying that they will maintain their normal shape (Gupta,  Ramsay, 1977). When this solution is hypotonic, water rushes into cells causing them to swell and burst if they are not strong (haemolysis for red blood cells). When the plasma is hypertonic (more concentrated than the cells), the cells lose water and shrink (Gupta,  Ramsay, 1977). 

Shrinking or bursting of cells due to osmotic and permeability relations of cells can have detrimental effects on their functioning (Limbeck, Arthur,  Nachbar, 1971).  It is therefore important for the plasma to remain in an isotonic state and for the cells to maintain their osmotic pressure so that they can maintain their shape and function normally. Based on this, the study seeks to demonstrate the significance of osmotichydrostatic pressure in maintenance of cellular integrity using erythrocytes as model cells. In order to achieve this and comprehensively cover the topic, the study will also determine the isotonic coefficient of red blood cells. The study also seeks to establish the relationship between
1) Lipid solubility of organic molecules and permeability and
2) Molecular weight of organic molecules and permeability.

The experiment will use blood serum from fish that has diploid cells and triploid cells. According to Hyndman, Kieffer,  Benfey (2003), triploid fish offer a convenient model for studying the physiological significance of cell volume. The cardiovascular system of triploid cells is critical because of the potential effects the cell volume has on erythrocyte circulation. Triploid erythrocytes are more likely to experience greater resistance than diploid cells passing across constrictive regions such as the cell membrane or the microvasculature. However, cellular compensations for example increased deformation of the membrane or reduced cytoplasmic viscosity are likely to enhance maintenance of resistance that is similar to that one of passage of diploid red blood cells for the triploid erythrocytes passage (Benfey, 1999). It is expected in this study that larger triploid erythrocytes will experience greater resistance as compared to diploid cells which will take more time to pass through the cell membrane. Diploid cells are also expected to cause more damage to the erythrocytes.

Restriction enzymes are produced naturally by bacteria and cut the DNA at specific sequences. The sequences that are recognized are usually inverted repeats (palindromic) and the cuts that are made on the DNA are double stranded symmetrical cuts. Cleavage produces either cohesive or blunt ends. The mode of action of these enzymes is that they hydrolyze the phosphodiester bonds at specific cleavage sites present in the recognition sequences in each sugar-phosphate backbone of the DNA strand (Black, 2005 Pierce, 2003 Barnum, 1998 Stryer, 2002).

In partial digestion, the restriction enzymes are allowed to act on the DNA sequences for only a limited time. The objective is to prevent the enzyme from cleaving all the available restriction sites in the DNA. This generates large overlapping fragments or contigs that can be cloned using suitable vectors. Separation of the DNA fragments on the basis of their size differences can thereafter be carried out using gel electrophoresis (Pierce, 2003).

This experiment sought to partially digest the pRcCMV plasmid using BamH1 by varying the reaction times. BamH1 is an example of a type II restriction enzyme and it is isolated from Bacillus amyloliquefaciens. The recognition sequence for this enzyme is 3-CGATCC-5 and it produces cohesive ends. The pRcCMV plasmid is 5542 bp long and has 3 BamH1 restriction sites and these are situated at 909 bp, 1306 bp and 3350 bp respectively (Pierce, 2003). Since the DNA is circular, cleavage of the plasmid using BamH1 was expected to yield 3 fragments for each of the test reactions. The sizes of the fragments were expected to be 397 bp (1306-909), 2044 bp (3350-1306) and 2192 bp (5542-3350) respectively. The control was expected to yield one fragment since no cleavage occurs as the plasmid DNA was not exposed to the enzyme. The band representing the fragment produced was expected to be located close to the loading point due to its relatively big size.

Materials and methods
The BamH1 restriction enzyme was used to cleave the plasmid DNA in reactions that were performed at 37c. The reactions were carried out in 4 different tubes, one of which was the control tube. The reaction buffer, the plasmid DNA and enzyme were added to all the test tubes. No enzyme was added to the control tube. The reactions in the 3 test tubes were timed and were allowed to proceed for 2, 10 and 30 minutes respectively. GlycerolEDTA was used to stop the reactions. The loading dye was added into each of the 4 tubes, the gel prepared and the samples loaded into separate wells. The gel was then placed into the tank and run for 5-7 minutes at 275V. The bands obtained were then photographed.

Difficulties encountered during the experiment included leakage of the gel mixture and that the gel had speckles after destaining had been done. Leakage of the gel mixture was stopped by correctly aligning the spacers and the bottom of the glass plates. To get rid of the speckles, the stain was filtered and this removed the precipitates present which were the cause of the speckles.