Laboratory Animal Science 48 (2)
1998

Brownstein, DG Genetically Engineered Mice: The Holes in the Sum of the Parts. Laboratory Animal Science 48 (2): 121.
This is an editorial which describes some of the issues which laboratory animal veterinarians should be concerned about when dealing with transgenic mice.  Suggests research needed into the peripheral effects of genetic engineering.  The author reflects on the less-emphasized aspects of transgenic mice, namely the reproductive biology, embryology and genetics.  With the onset of transgenics 18 years ago in a report by J.W. Gordon et al, an unprecedented share of biomedical research has relied on genetically engineered mice.  However, there is a scarcity of publications that deal directly with the mice, due to lack  of familiarity of the model and the technology (according to the author).

Areas that need further investigation include: methodology to increase fecundity,  assessing lineage to understand the consequences of mixing inbred mouse genomes, and assessing the consequences of allogeneic foster pregnancies
Questions:
1.  When and where was the first paper published on transgenic mice?
Answer:
1.  Gordon, J.W., G.A. Scangos, D.J. Plotkin, et al. 1980.  Genetic transformation of mouse embyros by microinjection of purified DNA.  Proc. Natl. Acaed. Sci. USA  77:7380-7384.

Primary and Opportunistic Infections in Retroviral Induced Immunosuppression in Nonhuman Primates (Meeting Report) (B). Laboratory Animal Science 48 (2): 123.
The 29th Annual Primate Pathology Seminar and Workshop in Boston, Mass. was hosted by the Registry of Comparative Pathology and the New England Regional Primate Research Center (Feb. 28-March 1).
      The majority of the presentations centered on the manifestations ofSIV (Simian immunodeficiency virus) infection but also included Simian retrovirus (SRV), Simian-human immunodeficiency virus (SHIV), and human immunodeficiency virus (HIV).  One speaker described "inflammatory conditions associated with SIV infection, including arteropathy/arteritis, multi-nucleate giant cell disease, and data associating chemokine and chemokine receptor expression wtih the genesis of encephalitic lesions."
Data was presented on AIDs in an HIV infected chimpanzee.
      Opportunistic infection of immunosuppressed macaques were described.  These included Pneumocystis carinii infection, mycobacteriosis, cryptosporidiosis, rhesus cytomegalovirus infection, adenovirus infection, and candidiasis.  Enterocytozoon bieneusi in immunocompromised macaques was discussed as related to the same pathogen which causes morbidity and mortality in human AIDs patients.
      "Potential nonhuman primate models of AIDs-associated neoplasia were described."  A herpesvirus of macaques may be related to human herpesvirus 8 (HHV-8) which may cause Kaposi's sarcoma, Castleman's diesease, and body cavity lymphoma.  An Epstein-Barr virus (EBV)-like virus related lymphosarcomas in SIV infected macaques as a potential model of
HIV-associated lymphomas in humans.
      Next year's meeting will precede the United States-Canadian Academy of Pathology in San Francisco.  For details call Dr. O'Neill at the Registry of Comparative Pathology, Armed Forces Institute of Pathology, Washington, D.C. 20306-0001.
      For more information on covered subjects, the following publications can be consulted:
     "Development of AIDS in a Chimpanzee Infected with Human
Immunodeficiency Virus Type I", Journal of Virology 71(5): 4086-4091,1997.
     "Identification of an Enterocytozoon bieneusi-Like Microsporidian Parasite in Simian-Immunodeficiency-Virus-Inoculated Macaques with Hepatobiliary Disease", American Journal of Pathology 150(4): 1395-1405, 1997.
     "A Herpesvirus of Rhesus Monkeys Related to the Human Kaposi's Sarcoma-Associated Herpesvirus", Journal of Virology 71(12): 9764-9769, 1997.
No Questions.

Casebolt, DB, DJ Speare, and DG Brownstein Care and Use of Fish as Laboratory Animals: Current State of Knowledge. Laboratory Animal Science 48 (2): 124.
This is an excellent article which is an overview of the use of fish in the laboratory setting.  Water quality, which must be defined based on the individual species, stage of life, and research, is the most important element of fish husbandry.  Effective environmental control is essential for effective disease control.  Stocking density and rate of water changes need to be considered together.  When evaluating health records of fish, keep in mind that certification for international trade does not imply that the fish are healthy - only states they are free from diseases important in international trade.  Excellent chart of differential diagnoses, but authors leave descriptions of diagnosis and treatment to the available textbooks.
Questions:
1)  Which of the following may have an effect on the physiological aspects of growth?
a.  Lighting intensity
b.  Noise levels
c.  Stocking density
d.  Feed delivery systems and timing of delivery
e.  All of the above.
2)  Water is important to fish because it:
a.  Provides their oxygen
b.  Provides their food
c.  Is thermoregulatory.
d.  All of the above.
3)  True of False:  The adequacy of flow rates (liters/minute) of water in a tank is generally viewed in the context of the upper concnetration of metabolic oxygen (mgo2/kg fish/min) demand of the tank inhabitants.
4)  List three factors which can cause stress to fish in laboratory settings.
5)  List three ways to decrease stress in fish in laboratory settings.
Answers:
1)  E
2)  D
3)  True.
4)  Handling, experimental manipulation, temperature fluctuations, water-quality alternations, food availability, environmental noise, human activity, stocking density, social pressure, viral infection, bacterial infection, or parasitic infection.
5)  Use anesthetics (like tricaine methanesulfonate), lower water temps during transport and handling, add NaCl to freshwater fish tanks (slow acclimation needed).

Use of polymerase chain reaction to diagnose a natural outbreak of mouse hepatitis virus infection in nude mice  (B). Laboratory Animal Science 48 (2): 137.
     MHV is a mouse-specific corona virus with many strains.  Natural infection with MHV in mature, immunocompetent animals is often subclinical but in nude mice it could be a cause of a fatal wasting disease.  MHV has the potential to interfere with in vitro and in vivo research.
      This study describes a natural outbreak of disease due to MHV infection in a colony of nude mice in which confirmation of the diagnosis was pursued via serologic testing, histologic examination, EM, and reverse transcriptase -PCR (RT-PCR).  In this outbreak, nude mice obtained from a commercial SPF source were maintained in a barrier facility. Mice were used in tumor implantation study (prostate carcinoma).  The tumor cells had previously been passaged in other nude mice in Texas.  Nude mice implanted with the tumor cells developed wasting and diarrhea. All sera tested were negative for antibodies to MHV and other agents.  Immunocompetent sentinels introduced into the room also tested negative.  Histopathologic findings provided a presumptive diagnosis with focal hepatic necrosis and a few syncitia, also large multinucleate cells were seen in colonic an nasal epithelium.   EM results of hepatic tissues  were equivocal.  The etiology was eventually confirmed by RT-PCR.  Virus was detected in one nude and one sentinel mouse.  No virus was detected in cell lines that had been injected into the mice, hence the source of outbreak was unknown.
      By detecting viral genome, PCR offers an advantage over serologic testing in situations in which an antibody response is unlikely, e.g infection in immunodeficient animals, in young animals, and infection of cell cultures.  It is also a rapid and highly sensitive technique.  Limitations of PCR include its inability to provide retrospective evidence of infection because it requires the virus to still be present in the specimen.  Thus the need for complementary conventional techniques still remains.
Questions:
1)  Which of the following clinical signs can be seen grossly in the livers of mice infected with MHV?
a.  Multiple pale spots
b.  Pitting of capsular surface
c.  Fibrinous adhesions to surrounding structures.
d.  A and C.
e.  All of the above.
2)  How does one eradicate MHV from a colony?
3)  True or false:  The BALB/c mouse would be the appropriate sentinel for a nude mouse colony given their similar genetic backgrounds and their susceptibility to diseases like MHV.
Answers:
1)  E
2)  Depopulation
3)  True

Maggio-Price, L and others Diminished Reproduction, Failure to Thrive and Altered Immunologic Function in a Colony of T-cell Receptor Transgenic Mice:  Possible Role of Citrobacter rodentium. Laboratory Animal Science 48 (2): 145.
A transgenic colony with known MHV infection experienced a significant increase in mortality.  Citrobacter rodentium and several Helicobacter spp. were identified.  The Helicobacter spp remained constant, even after normal mortality rates had been re-established, therefore the authors concluded that C. rodentium was the inciting agent.  The authors were able to use enrofloxacin to eliminate C. rodentium from affected animals while rederiving via Ceasarian section.  They saw decreased ascites production during the interval where C. rodentium was present in the colony.
Questions:
1)  Citrobacter rodentium is a _______ and is transmitted via ________.
Answers:
1)  Gram negative rod; fecal-oral

Spontaneous Cholangiofibrosis in Long-Evans Cinnamon Rats:  a Rodent Model of Wilson's Disease (B). Laboratory Animal Science 48 (2): 156.
The Long-Evans Cinnamon (LEC) rat is a rodent model of Wilson's disease.  Wilson's disease in humans is caused by a deletion of the ATP7B gene which is thought to affect copper transport. LEC rats lack the rat homolog of the gene and subsequently develop ceruloplasmin deficiency, hepatic copper accumulation, and hepatocellular injury.  Cholangiofibrosis occurs after cell injury.
    Fifty-four LEC rats were studied at various ages (5-113 weeks).  Liver lesions were graded by the extent of involvement of cholangiofibrosis.  Metal analysis for copper and iron content were made on liver samples.    A positive correlation was found between the grade of cholangiofibrosis and age of the rats.  Jaundice was observed earlier in females although cholangiofibrosis develops at the same age in both sexes. Cholangiofibrosis was evident in all rats by 27 weeks of age and the condition progressed with age.  No statistically significant correlations could be made between concentrations of copper or iron and histologic grades.  Hepatic accumulation of copper is thought to be critical to the proper differentiation of cell along a biliary lineage.  Treatment with copper-chelating agents such as penicillamine or trientine prevents the development of cholangiofibrosis.
Questions:
1.  What are the pathologic changes in the liver of LEC rats?
2.  A.  What 3 substances can be administered to rats to experimentally
        induce cholangiofibrosis?
    B.  Where is the preferential localization of furan?
3. What is the only inbred strain of rats that spontaneously develops
   cholangiofibrosis?
Questions:
1)  What compounds can be used to induce cholangiofibrosis is rats on a choline-free diet?
a.  Furan
b.  Butter yellow
c.  Ethionine
d.  Alcohol
e.  A, B, and C
f.  All of the above
Answers:
1.  Fatty change, aneuploidy, proliferation of sinusoidal and mononuclear
    cells, inflammatory infiltration of hepatic parenchyma, hepatocellular
    dropout, regeneration, and fibrosis
2.  A. 1. furan
       2. butter yellow
       3. ethionine with a choline-free diet
    B. right caudate lobe of the liver
3.  Long-Evans Cinnamon rat

Herbst, LH and others Turmorigenicity of Green Turtle Fibropapilloma-Derived Fibroblast Lines in Immunodeficient Mice. Laboratory Animal Science 48 (2): 162.
This study found that GTFP-derived fibroblast could not be distinguished from normal skin derived fibroblasts by morphology and growth characteristics in vitro.  This makes it possible to investigate the molecular basis of cellular proliferation characterization of green turtle fibropapilloma.
Questions:
1.  What is the genus and species of the green turtle?
2.  What is the proposed reason why tumors grew on the footpads and the pinna, but not on the flank?
3.  What current models exist for assaying tumorigenicity of lower vertebrate cells?
Answers:
1.   Chelonia mydas
2.  The decreased temperature of the extremities are more accommodating for supporting growth of cells from poikilothermic species.
3.  The anterior chamber of the eye of Rana pipiens has been used to culture Lucke renal adenocarcinoma cells and irradiated Anolis carolinensis have been used for tumorigenicity studies of cell lines from several lower vertebrate species.

Lyme Borreliosis in Laboratory Mice  (B). Laboratory Animal Science 48 (2): 168.
Lyme disease in humans and other species is caused by infection with Borrelia burgdorferi, a tick-borne spirochete. The disease course is usually characterized by acute symptoms, followed by spontaneous remission and episodic bouts of recurrence, although some patients may develop chronic unremitting disease. B. burgdorferi has a broad natural host range.  It readily infects rhesus macaques, dogs, rabbits, guinea pigs, gerbils, hamsters, rats and mice. Mice are the best characterized model system because they are economical, genetically and microbiologically defined,  susceptible to disease at all ages, consistently develop heart and joint disease when inoculated by syringe or tick-borne infection, and disease induction does not require immunomodulation. There is a strong genetically determined susceptibility to disease: C3H/He mice develop relatively severe heart and joint disease; BALB/c mice develop mild heart and mild joint disease; and C57BL/6 mice develop mild heart and joint disease. Genetically based susceptibility to disease is apparently not immune mediated, since it is also expressed in mice with severe combined immunodeficiency (SCID). The tick Ixodes scapularis (a.k.a. I. dammini) is the vector of most North American B. burgdorferi infections. Following inoculation of the spirochete arthritis begins to evolve as early as 4 days and carditis by 7-10 days. Disease-susceptible C3H mice develop grossly visible tibiotarsal periarticular edema by 2 wks. Spirochetes are present in relatively small numbers but can be readily visualized in target tissues, such as joints and heart, during the acute phase of infection with a variety of methods, including silver stains, immunohistochemistry, or in situ DNA hybridization. The disease-resolution phase, which occurs after 30 days, requires functional immunity, as SCID mice, lacking T and B cells, develop persistent active carditis and progressively severe arthritis , with intense synovial proliferation and pannus of joints.
     The host immune response is primarily humoral. Studies of cellular immune responses are hampered by a nonspecific B-cell mitogenic effect of B. burgdorferi and its outer surface lipoproteins. The normally disease-resistant C57BL/6 mice can be converted to full disease susceptibility with the single beige (Chediak-Higashi) mutation. Beige mice lack functional NK cells and granulocytes. Lyme borreliosis has many clinicopathologic features in common with the human disease. One of the differences is that the mouse model does not seem to exhibit neurologic, dermatologic, or chronic unremitting arthritic lesion.
Questions:
1)  What causes Lyme disease?
2)  Which strain develops severe heart and joint disease?
3)  Which strain develops severe heart, but mild joint disease?
4)  Which strain develops mild heart and joint disease?
Answers:
1)  Borrelia burgdorferi, a tick-borne spirochete
2)  The C3H/He strain
3)  The BALB/c strain
4)  The C57BL/6 strain
Questions:
1. List three reasons why mice are the best characterized model system of
Lyme disease.
2. What is the most common tick species, which is the vector of most North
American B. burgdorferi infections.
3. Name three methods which can be used to visualize the spirochete in
target tissues?
4. The normally disease-resistant C57BL/6 mice can be converted to full
Lyme disease susceptibility with what mutation?
Answers:
1. Mice are the best characterized model system because they are
economical, genetically and microbiologically defined, susceptible to
disease at all ages, consistently develop heart and joint disease when
inoculated by syringe or tick-borne infection, and disease induction does
not require immunomodulation.
2. The tick Ixodes scapularis (a.k.a. I. dammini) is the vector of most
North American B. burgdorferi infections.
3. Silver stains, immunohistochemistry, or in situ DNA hybridization.
4. The single beige (Chediak-Higashi) mutation. Beige mice lack functional
NK cells and granulocytes.

Litwak, KN, WT Cefalu, and JD Wagner Strptozotocin-Induced Diabetes Mellitus in Cynomolgus Monkeys:  Changes in Carbohydrate Metabolism, Skin Glycation, and Pancreatic Islets. Laboratory Animal Science 48 (2): 172.
Spontaneous adult-onset diabetes mellitus in NHP is an excellent model of human disease (both associated with increased age and obesity, changes in carbohydrate metabolism, variable changes in plasma lipids and lipoprotein concentrations), but the slow development and low frequency of occurrence limits its usefulness. Streptozotocin was used to induce diabetes.  Streptozotocin induces a state of chronic hyperglycemia due to its toxic effect on pancreatic beta cells. Chronic hyperglycemia leads to glycation of numerous proteins. Glycation products result in advanced glycated end products (AGE). These events may contribute to the chronic diseases associated with diabetes. The authors wanted to determine if this model could be used to study mechanisms of diabetes associated cardiovascular disease.
        Parameters followed included blood glucose, glucose tolerance, glycated hemoglobin levels, skin AGE levels, plasma total cholesterol, HDL-C, VLDL, LDL-C, and TG, and tissue histopathology. Two months following STZ, monkeys were glucose intolerant, had increased glucose, glycated hemoglobin, and skin AGE levels. Glycation of hemoglobin equilibrates with ambient glucose concentrations, whereas skin AGE content increases over time in both diabetic and non-diabetic subjects and is considered an index of aging. No change in any cholesterol levels was seen as long as glycemia was controlled, similar to humans with well controlled type-1 diabetes. No visible changes in islet structure were seen but insulin immunostaining decreased. This is expected in light of the fact that STZ causes dose-dependent damage and not autoimmune insulitis. By using a lower dosage of STZ the authors were able to induce a model of chronic hyperclycemia, void of negative hepatic and renal effects, with minimal changes in lipid or lipoprotein concentrations, and concluded that this would be a useful model for studying its relationship with cardiovascular disease.
Questions:
1. What other disease has STZ been used to study in NHP?
2. List 4 conditions induced by vascular changes associated with diabetes
mellitus and chronic hyperglycemia.
3. True or false: The percentage of islet insulin staining and physiologic
responses to the IV glucose tolerance tests showed  a high level of
correlation in all monkeys tested.
4. True or false: Insulin deficiency causes an increase in lipoprotein
lipase activation.
Answers:
1. Atherosclerosis
2. Nephropathy, retinopathy, neuropathy, and vasculopathy.
3. True
4. False

Hypoxanthine Phosphoribosyltransferase cDNA in Gerbils (Meriones unguiculatus). Laboratory Animal Science  48 (2): 179.
Hypoxanthine Phosphoribosyltransferase (HPRT) is a purine salvage enzyme that catalyzes conversion of hypoxanthine and guanine to their respective mononucleotides. Biosynthesis of purine and pyrimadine nucleotides is a crucial process in all growing cells because these molecules are the direct precursors of DNA and RNA. Constitutive expression of the mRNA for HPRT has been exploited for the use as an internal control in reverse transcriptase-polymerase chain reaction (RT-PCR) quantification of cytokine roduction because of its ubiquitous nature. Mammalian HPRT genes have been cloned and sequenced for the molecular analysis of Lesch-Nyhan and gouty arthritis caused by a deficiency of this enzyme. This paper describes the cloning and sequencing of gerbil HPRT cDNA and the discovery of two unique forms of the cDNA. Having these cloned genes available for the gerbil can be used to determine the pattern of cytokine gene expression in the gerbil (jird). Gerbils are used as models for a variety of diseases, including infection with Helicobacter pylori, Schistosoma mansoni,
Acanthocheilonema vitae and Brugia pahangi.
Questions:
1. Deficiency of the mammalian HPRT enzyme causes what two diseases?
2. List three diseases in which gerbils are used as models.
Answers:
1. Lesch-Nyhan and gouty arthritis are caused by a deficiency of this enzyme.
2. Helicobacter pylori, Schistosoma mansoni, Acanthocheilonema vitae and
Brugia pahangi

"Absence of  a Significant  Mixed Lymphocyte Reaction in a Marsupial (Monodelphis domestica)". Laboratory Animal Science 48 (2): 184.
In a previous study it  was suggested that the marsupial Monodelphis domestica failed to exhibit a mixed lymphocyte reaction (MLR) with allogeneic lymphocytes.  This study was to see if this was a matter of a response too weak to detect but capable of being augmented by immunization.  The results of this study plus other studies with other marsupials, strongly indicate that the failure to have a significant  MLC response is a general feature of marsupials.  This suggestion seems to fit with the accepted view that marsupials tend to have a more primitive immune response than do eutherian mammals.  It is generally accepted that the MLC response is a measure of T-cell function in concert with genetic polymorphism at the major histocompatibility (MHC) class-II loci.  Some studies suggest that a lack of polymorphism at the class-II MHC locus accounts for the lack of MLC response.  This study provides information that this is not the case in M. domestica.  It was shown that there is polymorphism at the class-II locus in M. domestica.  Further molecular studies are being conducted.
"It seems likely that the failure of this species to exhibit a significant  mixed lymphocyte response is due to T cells whose ontogeny differs from that of the T cells of eutherian mammals".
Questions:
1.  Define "kinship coefficients"
2.  The immune responses of M. domestica are very similar to what type of mouse?
3.  What is the genus a species of    a. koala b. tammar wallaby
Answers:
1.  A measure of the genetic relatedness between two animals
2.  a mouse lacking alpha/beta T cells due to a targeted mutation in the T-cell receptor at the  beta locus
3.  a.  Phascolarctos cinereus
b.  Macropus eugenni

Diagnostic Exercise:  Lethal Pneumonia in Neonatal Kittens. Laboratory Animal Science 48 (2): 190.
 Approximately 2 weeks after birth, a litter of two kittens born to a primipara queen used in a fetal alcohol syndrome study developed acute respiratory distress and died within 24 - 48 hours.  Gross necropsy findings were congested and consolidated cranial lung lobes. Histopathology of this tissue revealed that the bronchial and bronchiolar epithelium was necrotic, the lumens contained variable amounts of cellular debris, fibrin and neutrophils.  There was multifocal patchy necrosis of alveoli, with flooding of lumens with fibrin and cellular debris.  Nuclei of airway epithelial cells contained eosinophilic inclusions surrounded by a rim of marginated chromatin.
 The diagnosis was infection with feline herpes virus, which can rarely cause severe fatal pulmonary disease characterized by necrotizing bronchitis, bronchiolitis, and pneumonia, with serofibrinous flooding of alveoli and airways in young kittens.  In this case the mother was likely a carrier that had a recrudescence and viral shedding because of the stress of pregnancy and in turn infected her young kittens.
 Questions:
1.  List 6 differential diagnosis for feline pneumonia.
2.  What are 2  metazoan parasites that can infect the feline lung?
Answers:
1.  viral,  bacterial (including Chlamydia) ,  fungal,  protozoan parasites, metazoan parasites, foreign body/aspiration.
2.   Aelurostrongylus obstrusus and Paragonimus kellicotti

Further Evaluation of a Diagnostic Polymerase Chain Reaction Assay for Pasteurella pneumotropica. Laboratory Animal Science 48 (2): 193.
 This article summarizes the evaluation of PCR for Pasteurella pneumotropica by using primers PPN-1 and PPN-2.  Test DNA had been extracted from bacterial suspensions via boiling, precipitation of lysoszyme and proteinase K digests, via CTAB/NaCl purification methods followed by RNAse I digestion and reprecipitation.  All P. pneumotropica reference strains and field isolates were identified by PCR; however, Actinobacillus muris also had positive  results.  A. muris disappeared when the annealing step was increased from 55 deg. C to 57 deg. C.  A. muris has been isolated from conjunctiva, oropharynx, and lower respiratory tracts of mice, with or without evidence of inflammation.  The use of PCR may definitively identify Pasteurella pneumotropica and differentiate it from Actinobacillus muris.
QUESTIONS:
1.  Pasteurella pneumotropica is a gram-negative coccobacillus harbored
by:
        a. laboratory rats
        b. mice
        c. hamsters
        d. guinea pigs
        e. other rodents
        f. a., b., & c.
        g. a. thru e.
2.  PCR assays have higher specificity than do traditional diagnostic methods and save technician time and large amounts of biochemical reagents required to distinguish it from related bacteria of which complex:
        a. Haemophilus-Pasteurella-Streptococcus
        b. Haemophilus-Pasteurella-Bordatella
        c. Haemophilus-Pasteurella-Actinobacillus
        d. Streptococcus-Pasteurella-Actinobacillus
        d. Streptococcus-Pasteurella-Bordatella
3.  Define the Acronyms:
        a. PCR
        b. RAPD-PCR
        c. CTAB
        d. NIEHS
        e. ATCC
4.  By varying components of the PCR assay and keeping other factors constant, cross-reactivity with Actinobacillus muris  disappeared when the
        a. Annealing step was increased from 55 deg. C to 60 deg. C.
        b. Denaturation step was decreased from 94 deg. C. to  90 deg C.
        c. Extension step was increased from 72 deg C. to 75 deg C.
        d. Denaturation step was increased from 94 deg. C. to 98 deg. C.
        e. Annealing step was increased from 55 deg. C. to 57 deg. C.
ANSWERS:
1.  Pasteurella pneumotropica  is a gram-negative coccobacillus harbored by:
        g. laboratory rats, mice, hamsters, guinea pigs, and other rodents.
2.  PCR assays have higher specificity than do traditional diagnostic methods and save technician time and large amounts of biochemical reagents required to distinguish it from related bacteria of which complex:
       c. Haemophilus-Pasteurella-Actinobacillus
3.  Define the Acronyms:
        a. PCR          Polymerase Chain Reaction
        b. RAPD-PCR     Randomly Amplified Polymorphic DNA - Polymerase Chain
        c. CTAB         Cetyltrimethylammonium Bromide
        d. NIEHS        National Institute of Environmental Health Sciences
        e. ATCC         American Type Culture Collection
4.  By varying components of the PCR assay and keeping other factors constant, cross-reactivity with Actinobacillus muris  disappeared when the
       e. Annealing step was increased from 55 deg. C. to 57 deg. C.

Comparison of Bile Chemistry Between Humans, Baboons, and Pigs: Implications for Clinical and Experimental Liver Xenotransplantation. Laboratory Animal Science 48 (2): 197.
 Despite the immunologically encouraging results of two baboon-to-human liver transplants, both of the grafted baboon livers
developed cholestasis (sludging) in the common bile duct. Pig liver is rejected rapidly in non-human primates with a maximal survival of only 3 days.  With the emphasis on developing a successful porcine xenotransplant model, the etiology of  ôsludgingö from baboon-human model must be elucidated.  This paper examines gallbladder and hepatic bile of humans, baboons, and pigs to determine viscosity, pH, bilirubin, cholesterol, and electrolyte concentrations.
        No significant differences in gallbladder viscosity, pH, cholesterol, Na, Cl, HCO3, and P concentrations in the pig, baboon, and human.
QUESTIONS:
1.  How was viscosity measured?
2.  In gallbladder bile  mean K, Ca, and bilirubin concentrations were (higher, lower) (chose one) than in baboons, and mean Ca, Mg, and total and indirect bilirubin were (higher, lower) (chose one) in humans than in pigs.  Cholyglycine values varied widely within each species but were highest in (humans, baboons, pigs) (chose one).
3. In liver bile, viscosity, cholesterol, Na, K, Ca, Mg, and total bilirubin were significantly (higher, lower) (chose one) in humans than in (baboons, pigs) (chose one).
4. In liver bile, cholyglycine values were (lower, higher) (chose one) in baboons compared to humans and pigs.
5. Both grafted baboon livers in clinical xenotransplantation contained inspissated bile ("sludge") in the (gallbadder, bile duct, extrahepatic tissues) (chose one).
6. If you received a liver transplant from a baboon, would you also receive the baboon's gallbladder?
7. Were there any differences found in gallbladder or hepatic bile which would account for the sludging of bile produced by baboon or pig livers in humans or by pig livers in baboon?  What might account for the sludging?
ANSWERS:
1.  How was viscosity measured?
        Capillary Flow viscometer
2.  In gallbladder bile  mean K, Ca, and bilirubin concentrations were higher than in baboons, and mean Ca, Mg, and total and indirect bilirubin were higher in humans than in pigs.  Cholyglycine values varied widely within each species but were highest in baboons.
3. In liver bile, viscosity, cholesterol, Na, K, Ca, Mg, and total bilirubin were significantly higher in humans than in baboons.
4. In liver bile, cholyglycine values were higher in baboons compared to humans and pigs.
5. Both grafted baboon livers in clinical xenotransplantation contained inspissated bile ("sludge") in the bile duct.
6. If you received a liver transplant from a baboon, would you also receive the baboon's gallbladder?  No, because a cholecystotomy is performed during the transplantation.
7. Were there any differences found in gallbladder or hepatic bile which would account for the sludging of bile produced by baboon or pig livers in humans or by pig livers in baboon?  No  What might account for the sludging? Human environment alter the character of bile after xenotransplantation, or immunologic factors may play a role.

Modified Technique for Abdominal Heterotopic Cardiac Transplantation in the Rabbit. Laboratory Animal Science  48 (2): 201.
Heterotopic cardiac transplantation has been used to evaluate complications of transplant procedures, evaluate new drugs, and study the pathogenesis of graft survival.  Techniques for abdominal heterotopic cardiac transplant have been described including end-end anastomosis (donor aorta and pulmonary artery to recipient abdominal aorta and CVC at the level of the renal vessels) which resulted in paraplegia and paralysis and an improved end-side anastomosis.  The end-side anastomosis resulted in 100% incidence of paralysis in rabbits weighing > 1 kg.  This article describes a modified cross clamp technique that resulted in no signs of spinal cord ischemia in the 41 rabbits used during this study.  The improved cross clamp technique makes the abdominal heterotopic cardiac transplant a valuable and reliable tool for further studies in transplantation.
No questions

Doppler Echocardiographic Analysis of Effects of Tribromoethanol Anesthesia on Cardiac Function in the Mouse Embryo:  A Comparison with Pentobarbital. Laboratory Animal Science 48 (2): 206.
This article compared doppler echocardiography in pregnant female mice anesthetized with either tribromoethanol (TBE) or pentobarbital sodium (PB).  TBE induced transient arrhythmia in mouse embryo hearts mainly between days 11 and 13 of gestation.  This suggests that the mouse embryo heart is especially vulnerable to the anesthetic during this period.  There was no arrhythmia in the PB group of embryos.  The heart rate in the TBE group was slower than in the PB group. (TBE) has been an extensively used anesthetic in research involving mice because of its rapid onset of effect, wide range of tolerance, and low mortality. They concluded that TBE can induce arrhythmias and/or can inhibit the conduction at these developmental stages.
Questions:
1.      What anesthetic induced arryhthmias in mouse embryo hearts?
2.      TBE induced transient arrhythmias in mouse embryo hearts between days
        a.1-3
        b. 4-6
        c. 7-9
        d. 11-13
Answers not provided.

Detection of Mycoplasma pulmonis from Rats and Mice of Sao Paulo/SP, Brazil. Laboratory Animal Science 48 (2): 210.
Mycoplasma infection of the respiratory tract is a major disease entitiy in laboratory rats and mice.  As well, Mycoplasma can cause joint and vaginal infections.  Mycoplasma infection can cause significant interference with experimental data and lead to potential misinterpretation of the data.
    One barrier-maintained facility was found to have no mycoplasma (by culture of tracheobronchial lavage or ear flushing samples); however all of the conventional facilites were found to have a high positive culture rate.  Tracheobronchial lavage was a more reliable technique.  Facility refurbishment and hygiene controls failed to eliminate M. pulmonis infection in 3 facilties.
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