three autosomal genes (Z, E and T) in seahorses. Each gene sorts independently and each gene has two alleles. The Z allele forms zebra stripes which are completely dominant over no stripes, z. The E allele codes for crossed eyes which is dominant over uncrossed eyes, e. Lastly, the T allele codes for jellyfish-like tentacles,which are dominant over (t) squid-like tentacles. You have two parents that are completely heterozygous for all traits. What is the probability they have offspring that have:

a.Zebra Stripes, uncrossed eyes, and squid-like tentacles? ________________

b.All dominant traits?_______________________________

c.Have the genotype ZZeeTt?_________________________

d.True breeding for zebra stripes, crossed eyes and squid-like tentacles?______________

e.All dominant alleles or all recessive alleles?________________

f.The first progeny is a male with zebra stripes and the second progeny is a male with no stripes?(assume sex probabilities in zebrafish is similar to humans)

Which of the following is a typical characteristic of human traits that follow an X-linked recessive inheritance pattern?

Group of answer choices

more males than females are affected

more females than males are affected

it is passed from father to all sons

affected sons must have an affected mother

True or False - Du positive blood is regarded as Rh-negative for transfusion purposes.

Why is it preferable, if possible, to take a narrow spectrum antibiotic over a broad spectrum antibiotic?

Group of answer choices

narrow spectrum antibiotics have a limited range of effectiveness among different bacterial species but they each have several modes of action which make them more effective than broad spectrum antibiotics

narrow spectrum antibiotics are less likely to be overcome by antimicrobial resistance mechanisms compared to broad spectrum antibiotics

narrow spectrum antibiotics always have a high theraputic index compared to broad spectrum antibiotics

narrow spectrum antibiotics are less likely to destroy the normal gut microflora which can lead to development of super-infections after treatment

narrow spectrum antibiotics attack many more species of bacteria compared to broad spectrum antibiotics

Using the Arizona Wuhan seafood market virus isolate, MN997409.1, run BLAST against the RefSeq Representative genomes (refseq_representative_genomes) database, limiting the search to Viruses (taxid: 10239) in the “Organism” field. From the hits returned, construct a phylogenetic tree (Distance tree of results). Examining the BLAST results and the Distance Tree select all that apply:

a)The non-Wuhan virus genome with the lowest E-value and highest identity is the SARS coronavirus

b)The distance tree suggests the Arizona virus is very different from the reference Wuhan seafood market virus isolate

c)The Distance tree suggests that the Wuhan viruses show similarity to a Bat coronavirus – BM48-31/BGR/2008

d)The White-eye corona virus show a significant hit, but with very low Query coverage

1)Vitamin A and pro-vitamin A are molecules that contain only carbon and hydrogen atoms and do not have any amino acids. If scientists want to genetically engineer a plant to make pro-vitamin A, what type of gene or genes would the scientists need to insert in the plant cells? Would they insert a gene that codes for pro-vitamin A? If not, what type of molecule would the gene or genes have to code for? Explain your reasoning.

2)Scientists have identified genes for two enzymes needed to make pro-vitamin A. One of these genes comes from corn. If this gene from a corn plant is inserted in the DNA of a rice plant, will the sequence of amino acids in the protein produced by the rice plant be the same as the sequence of amino acids in the protein produced by the corn plant? In other words, will rice plants that have this gene produce the same enzyme as corn plants produce? Explain why or why not.

3)Once scientists have identified the genes for enzymes to produce provitamin A, how could they insert these genes in the DNA of rice plant cells? Suggest one possibility.

4)Would you recommend that scientists try to insert the genes for enzymes to produce pro-vitamin A into:

a. all the cells in a rice plant

b. the thousands of cells in each rice grain or

c. a small group of embryonic rice plant cells that can divide and develop into a rice plant?

Explain your reasoning.

5) To insert genes from one organism into a different organism, scientists often take advantage of the natural genetic engineering capabilities of bacteria or viruses. One type of bacteria genetically engineers plant cells by inserting part of its bacterial DNA into the plant cell DNA, thus producing recombinant DNA. The inserted bacterial genes code for proteins that:

• stimulate the genetically engineered plant cells to produce food molecules that only the bacteria can use
• stimulate these genetically engineered plant cells to divide and form a growth that bulges out from the stem or root.

Explain how this type of genetic engineering is useful for the bacterium.

fluid moving into the brain after a traumatic brain injury can result in swelling of the brain. what type of solution(hypertonic, isotonic, hypotonic)would doctors use to relieve the swelling. why. 5 marks

Define multihit hypothesis. A complete definition will include the following (4pts):

·Define the term “hit”.(1)

Accumulation of mutations in genes that control cell cycle, apoptosis, and cell migration in a adult stem cell.

·Define how many hits normally will cause a cell to become cancer.(1)

·Explain the type of cell that normally becomes a cancer cell and why.(1)

·Finally, make sure to explain the importance of time in a cell becoming a cancer cell.(1)

Question 21

1. Vibrios are bacteria that have.....

   	A.	Comma shape
   	B.	Rod shape
   	C.	All are correct
   	D.	Spiral shape

2.5 points

Question 22

1. Fungal hyphae in which two genetically distinct kinds of……………………occur together are said to be heterokaryotic

   	A.	Nuclei
   	B.	Chloroplast
   	C.	Parenchyma cells
   	D.	Flagella

2.5 points

Question 23

1. ............................occurs in vascular plants which have microspores and megaspores

   	A.	Algae
   	B.	Diatom
   	C.	Heterospory
   	D.	Bacteria

2.5 points

Question 24

1. The first plants clearly evolved from an organism that, if it existed today, would be classified as a multicellular…………………

   	A.	Cyanobacteria
   	B.	Bryophyte
   	C.	Green algae
   	D.	Meristematic tissue

2.5 points

Question 25

1. Chloroplasts are descended from

   	A.	Plasmid
   	B.	Fern frond
   	C.	Cyanobacteria
   	D.	Trypanosoma

2.5 points

Question 26

1. Shigella species causes

   	A.	Measles
   	B.	Pneumonia
   	C.	Bacterial descentry
   	D.	Chicken pox

2.5 points

Question 27

1. Which of the following statement is not true about protists?

   	A.	Protists have cuticle
   	B.	Photosynthetic protists are at the base of aquatic food chain
   	C.	Protozoan are the heterotropic unicellular protist
   	D.	Euglena is a photosynthetic protist

2.5 points

Question 28

1. Humankind's most prevalent diseases are caused by phylum apicomplexan which are protists consist of endoplasmic protozoan. Which of the following disease is caused by these protists?

   	A.	Head injury
   	B.	Malaria (by Plasmodium)
   	C.	Fungal infection
   	D.	Any physical trauma

2.5 points

Question 29

1. Which of the following is true regarding photosynthetic protists?

   	A.	They are called lichens
   	B.	They are called photoautotroph
   	C.	They are parasites
   	D.	They do genetic transduction

2.5 points

Question 30

1. What does Lichen mean?

   	A.	Helpful symbiotic relationship between blue/green algae (cyanobacteria)  with filamentous fungus
   	B.	It is an insect
   	C.	It is harmful for mankind
   	D.	All are correct

1a. If a 10^-2 dilution of a culture yields 150 colonies when 0.5 ml is spread on a plate, how many colonies will grow on a plate that receives 0.1 ml of the same dilution?

1b. How many CFU per ml are present in a sample if 0.2 ml of a 10^-5 dilution grew 220 colonies?

1c. How many colonies do you expect when 0.4 ml of a 10^-7 dilution of a culture containing 10^9 CFU per ml is plated onto nutrient media?

Complete the following dihybrid crosses. Draw out the Punnett for the F₁ generation and the F₂ generation.

1. In Drosophila, fruit flies, individuals with ebony body color and vestigial (small) wings represent the homozygous recessive. If you crossed an individual that was homozygous dominant (normal) for both genes with an individual that was homozygous recessive for both genes, you would observe the following:

a. phenotypic ratio in the F₁

b. phenotypic ratio in the F₂

c. genotypic ratio in the F₁

d. genotypic ratio in the F₂

After being exported to the cytoplasm, the 7-methyl guanosine cap was inadvertently (by mistake) cleaved off of a eukaryotic mRNA. As a consequence, which of the following will probably happen?

Read the following and answer the questions.

INTRODUCTION

Oral cavity is an open growth system with an uninterrupted introduction and removal of microbes and their nutrients. It offers diverse habitats where-in different species of micro-organisms can prosper. The primary requisite for any group of microbes to flourish in a niche is their ability to adhere to the tooth surfaces and multiply in shielded environments like periodontal pockets and tooth crevices. Such an aggregation of microbes on tooth surfaces has been traditionally referred to as ‘plaque’ because of its yellowish color, reminiscent of mucosal plaques caused by syphilis.

Dental plaque has been defined as “a specific but highly variable structural entity consisting of micro-organisms and their products embedded in a highly organized intercellular matrix.” It represents a true biofilm consisting of a variety of micro-organisms involved in a wide range of physical, metabolic and molecular interactions. The cooperative nature of a microbial community provides advantages to the participating organisms such as a broader habitat range for growth, enhanced resistance to antimicrobial agents and host defenses and enhanced pathogenicity.[1]

Biofilms have been implicated as the chief culprit in the etiopathogenesis of dental caries and periodontal disease. Though uncalcified biofilms can be removed by routine oral hygiene aids or professional dental instruments, they have the potential to calcify into dental calculus making their removal difficult. Hence, these biofilms pose a great challenge to the dental clinician in the control and eradication of biofilm-associated diseases.

HISTORICAL PERSPECTIVE

Biofilms are nothing new. The first description dates back to the 17^thcentury, when Anton Von Leeuwenhoek - the inventor of the Microscope, saw microbial aggregates (now known to be Biofilms) on scrapings of plaque from his teeth.

The term ‘Biofilm’ was coined by Bill Costerton in 1978.

In 2002, Donlan and Costerton offered the most salient description of a biofilm. They stated that biofilm is “a microbially derived sessile community characterized by cells that are irreversibly attached to a substratum or interface or to each other, embedded in a matrix of extracellular polymeric substances that they have produced, and exhibit an altered phenotype with respect to growth rate and gene transcription.”[2]

WHAT IS A BIOFILM?

The term Biofilm (Wilderer and Charaklis 1989) describes the relatively indefinable microbial community associated with a tooth surface or any other hard non-shedding material, randomly distributed in a shaped matrix or glycocalyx.[2]

In the lower layers of a biofilm, microbes are bound together in a polysaccharide matrix with other organic and inorganic materials. Above it, is a loose amorphous layer extending into the surrounding medium. The fluid layer bordering the biofilm has stationary and dynamic sub layers.

FORMATION OF A BIOFILM

Formation of a biofilm is a complex process that follows several distinct phases, beginning with adsorption on to the tooth surface of a conditioning film derived from bacterial and host molecules, which forms immediately following tooth eruption or tooth cleaning. This adsorption is followed by passive transport of bacteria mediated by weak long-range forces of attraction. Covalent and hydrogen bonds create strong, short-range forces that result in irreversible attachment.

The primary colonizers form a biofilm by autoaggregation (attraction between same species) and coaggregation (attraction between different species). Coaggregation[4] results in a functional organization of plaque bacteria and formation of different morphologic structures such as Corncobs and Rosettes. The microenvironment now changes from aerobic/capnophilic to facultative anaerobic. The attached bacteria multiply and secrete an extracellular matrix, which results in a mature mixed-population biofilm.

After one day, the term Biofilm is fully deserved because organization takes place within it. Transmission occurs from other sites, leading to incorporation of new members into the biofilm and the formation of a climax community. The thickness of the plaque increases slowly with time, increasing to 20 to 30 μm after three days.

Four stages of dental plaque biofilm growth (as shown in Figure 1)

Stage I - Attachment (lag - not inert, but metabolically reduced)

Stage II - Growth (log - exponential growth)

Stage III - Maturity (stationary)

Stage IV - Dispersal (death)

PROPERTIES OF BIOFILMS

Biofilms are ubiquitous; they form on virtually all surfaces immersed in natural aqueous environments. A biofilm confers certain properties to bacteria that are not seen in the planktonic state, a fact that justifies recognition of dental plaque as a biofilm.

A major advantage is the protection that biofilm provides to the colonizing species from competing micro-organisms, environmental factors such as host defense mechanisms and potentially toxic substances like lethal chemicals or antibiotics. Biofilms also facilitate processing and uptake of nutrients, cross feeding and removal of potentially harmful metabolic products through the voids or water channels between the micro-colonies, acting as a primitive circulatory system.[1] They also create an appropriate physicochemical environment such as a properly reduced oxidation reduction potential.

An important characteristic seen in Biofilm-associated bacteria is Quorum sensing, or cell density mediated gene expression.[5] This involves the regulation of expression of specific genes through the accumulation of signaling compounds that mediate intercellular communication. Quorum sensing may give biofilms their distinct properties. Eg.- Expression of genes for antibiotic resistance at high cell densities may provide protection. It also has the potential to influence community structure by encouraging the growth of species beneficial to the biofilm and discouraging the growth of competitors.

Another important characteristic of biofilm associated bacteria is the gene transfer[6] through which bacteria communicates with each other. In S. mutans, quorum sensing is mediated by competence stimulating peptide, wherein genes are responsible for multiple functions - biofilm formation, competence and acid tolerance.

Biofilm related Regulation of gene expression has been shown in certain bacteria. eg. Exposure of S. gordonii to saliva results in induction of genes that mediate host surface binding and coaggregation with P. gingivalis and Actinomyces. Similarly, genes encoding glucan and fructan synthesis are differentially regulated in Biofilm-associated S. mutans.

MECHANISM OF INCREASED ANTIBIOTIC RESISTANCE IN BIOFILMS

Organisms in a Biofilm are 1000-1500 times more resistant to antibiotics than in their planktonic state. The mechanisms[2] of this increased resistance differ from species to species, antibiotic to antibiotic and for biofilms growing in different habitats. This antibiotic resistance in bacteria is thought to be affected by their nutritional status, growth rate, temperature, pH and prior exposure to sub-effective concentrations of antimicrobial agents. Another important mechanism appears to be the slower rate of growth of bacterial species in a biofilm, which makes them less susceptible to bactericidal antibiotics. Biofilm matrix can resist diffusion of antibiotics. Eg. strongly charged or chemically highly reactive agents can fail to reach the deeper zones of the biofilm as it acts as an ion exchanger in removing such molecules from solution.

‘Super-resistant’ bacteria have been identified within a biofilm, which have multidrug-resistance pumps that can extrude antimicrobial agents from the cell. Since these pumps place the antibiotics outside the outer membrane, the process offers protection against antibiotics that target cell wall synthesis. Above mentioned observations are critical to the use of antimicrobials in the treatment of Biofilm-associated infections.[7]

BIOFILMS AND INFECTIOUS DISEASES

Biofilms have been found to be involved in a wide variety of microbial infections (by one estimate 80% of all infections). These include dental caries, periodontal disease, otitis media, musculoskeletal infections, necrotizing fascitis, biliary tract infection, osteomyelitis, bacterial prostatitis, native valve endocarditis, meloidosis, cystic fibrosis pneumonia and peri-implantitis. Salient features of these infections are persistence and chronicity.[2]

What is biofilm and why can it be so dangerous?  

What is quorum sensing?  Discuss quorum sensing in terms of biofilm development?  

Explain the phrase, “dentistry focuses on biofilm.”