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Laboratory Animals Volume 44, Number 2, April 2010 original articles


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Laboratory Animals


Volume 44, Number 2, April 2010


ORIGINAL ARTICLES



Lourbopoulos et al. Predictable ventricular shift after focal cerebral ischemia in rats: practical considerations for intraventricular therapeutic interventions, pp. 71-78
Domain 1 (T1), Domain 3 (T3)

Primary Species: Rat (Rattus norvegicus)

 

SUMMARY: Transient infarction of the middle cerebral artery (t-MCAO) for various time periods in rats is commonly performed to model stroke.  The authors state that only 19.4% of animals can be injected properly using classical stereotaxic coordinates that do not take into account post infarction swelling and inflammation.  The authors performed the experiment to identify if there are predictable and consistent shifts in stereotaxic coordinates of the intracerebroventricular (ICV)space that occur in the 6-18 h period following t-MCAO.  The authors induced t-MCAO in Wistar rats for 2 hours, and evaluated animals using the modified Neurological Stroke Scale (mNSS), the modified Bedersons’ Scale (mBS), and the grid walking test (GWT) to determine if these scores could be correlated with infarction volume, hemispheric oedema, middle line dislocation and ventricular stereotaxic coordinates.  Following evaluations, a second stereotaxic surgery was performed to inject trypan blue to mark lateral ventricles using classical stereotaxic coordinates.  Brains were examined histologically to determine % hemispheric infarction volume, % oedema of the infracted hemisphere, slide diameter distance and depths, % middle line dislocation, and optimal site for stereotaxic invasion in each lateral ventricle.  They concluded that the brain ventricular system was displaced at AP levels of -0.12 to -2.04 from bregma.  Evaluation of the findings determined two numerical constants, fixed number and factor, b0 and b1 respectively, by regression analysis, that in 80% of animals could be used to predict accurate new stereotaxic coordinates for ICV injection that are shifted by post infarction swelling and inflammation.  The following equation is proposed for use to verify appropriate new coordinates for each animal – stereotaxic coordinate =b0 +b1*mNSS.    While mNSS was the only factor used, the authors state that mBS and GWT were also effective in predicting shift, but that mNSS is the most widely accepted stroke scoring method. The authors also note that MRI revealed that oedema was predictably present by 1 hour post, peaking at 24-48 hours. The authors state that it would be necessary to use @ 51 animals to successfully inject 10 under classical methods, and only 13 using the new methodology. It should be noted that animals were anesthetized with halothane in nitrous oxide and mechanically ventilated.  As halothane is no longer available in the US, for American colleagues, it may be necessary to revalidate the equation under isoflurane.

 

QUESTIONS:



  1. Name 3 standard methods of evaluating animals post t-MCAO for severity of stroke effect.

  2. What is the least predictable estimate of stroke damage of the 3 methods in question 1?

  3. How much is the brain shifted negatively or positively by edema post t-MCAO in the AP plane from bregma? 

  4. What is the rate of success of ICV injection using the modified coordinates? 

  1. 20% 

  2. 50% 

  3. 100% 

  4. 80%

ANSWERS:


1.    Modified neurological stroke score, modified Bedersons scale, and grid walking test.

2.    GWT

3.    Negative

4.    d. 80%

 

 

Diesch et al. Developmental changes in the electroencephalogram and responses to a noxious stimulus in anaesthetized tammar wallaby joeys (Macropus eugenii eugenii), pp. 79-87


Domain 2, Task 1

Tertiary Species: tammar wallaby joeys



SUMMARY: The age at which animals develop key neurological features in which they can perceive pain vary widely between species. Data obtained from rat pups (neurologically immature at birth) and sheep (neurologically mature at birth) demonstrates how different two species can be. However, that data cannot be extrapolated to marsupials which are born extremely immature and most of the neurological development occurs in the maternal pouch. The tammar wallaby joey (Macropus eugenii) is born at 28 days gestation with a cerebral cortex comprising 2 layers of cells and resembling that of a 40-day human embryo. It is not known when neurological development is sufficient to permit conscious sensory perception. The current study assessed when cerebral cortical processing of noxious sensory input may begin in the tammar wallaby joey using an approach previously used in rat pups and lambs, which evaluates EEG responses to potentially painful stimulation (toe clamping) in anaesthetized animals. The investigators found that the onset of sustained EEG activity followed pelage development and eye opening at 120-140 days of age. There was a continuous increase in power over all EEG frequencies at 180-190 days of age. Joeys aged 187-261 days exhibited a biphasic response to the noxious stimulus. The results do not indicate directly whether tammar wallaby joeys are able to experience pain but provide evidence of changes in cerebrocortical responsiveness to noxious stimuli. The greater cerebrocortical responses to noxious stimuli in older joeys and increased behavioural repertoire suggest tammar and other marsupial joeys are capable of conscious perception and pain experience.

QUESTIONS:



  1. Name 2 key neurological characteristics.

  2. What are 2 key markers of brain development that indicate joeys may consciously perceive noxious sensations?

  3. T/F. The sequence of EEG development is similar between mammals and marsupials but the timing differs significantly.

ANSWERS:


  1. 1) Animal must be sentient and

2) Brain must be functionally capable of inducing consciousness

  1. 1) opening of the eyes and

2) Development of the pelage

  1. T. Marsupials are much slower



Castelhano-Carolos et al. Identification methods in newborn C57BL/6 mice: a developmental and behavioural evaluation, pp. 88-103
SUMMARY: Individual identification of mice is important for a number of reasons and permanently identifying them at a young age after birth is necessary in a number of situations. In recent years, toe-clipping, a popular identification method of neonate rodents in the past, has been frowned upon. Many recommendations and guidelines suggest it is a method only acceptable when no others are feasible. The authors set out to investigate and compare the consequences of performing toe-clipping with another two permanent identification methods - toe tattoo ink puncture and s/c implantation of a small transponder - on 5-day old C57BL/6 mouse pups.
A total of 78 pups, from 17 different litters, were assigned randomly to each experimental group. Following permanent identification by one of the three methods listed above (a control group only has a s/c puncture), the pups were monitored immediately after and throughout development in the following ways: behavior at the time of identification (e.g., sudden movements, presence or absence of vocalizations, urination, etc); somatic parameters (e.g., incisors eruption, anogenital distance, opening of eyes, etc); neurological reflexes; locomotive coordination and muscular strength; grasping; surface righting reflex; postural reflex; bar holding test; postural stability and mobility test; air righting reflex. In addition, when the animals reached adulthood (from weaning until 15 weeks of age), they were assessed in the following ways: body weight; home-cage behavior; climbing; elevated plus maze; simplified SHIRPA screen; rotarod.
The authors found that all three methods were effective for long-term marking of mouse pups. In their assessment, newborn mice showed the greatest reaction/distress to the transponder implantation and the least to toe clipping. There were no overall differences in somatic and neurological reflex development during the postnatal period between groups and controls. Moreover, no adverse influences were found in general behavior or sensory-motor functions in adulthood. The authors conclude that toe clipping might even be advisable in newborn mice at a very young age as the best method of identification when genotyping is also required. In their study, it was the easiest and fastest method to perform, and the one that appeared to cause the least pain/distress in the animals.
QUESTIONS:

  1. At what age is it best to perform toe-clipping in newborn mice, when toes are large enough but ossification is not yet complete?

        1. Postnatal days 2-3

        2. After postnatal day 7

        3. Postnatal days 5-7

        4. Postnatal days 3-5

  2. The new, smaller transponders used in this study for implantation in newborn mice were:

        1. 2 mm in diameter and 12mm long

        2. 1 mm in diameter and 6mm long

        3. 2 mm in diameter and 4mm long

        4. 1 mm in diameter and 10mm long

  3. The Elevated Plus Maze test is used for assessing:

        1. Anxiety

        2. Posture

        3. Dominance

        4. Ambulation

  4. The Rotarod test is normally used for assessing:

        1. Posture

        2. Gripping ability

        3. Reflexes

        4. Motor coordination and balance

  5. True or False: Tissue obtained from toe-clipping of mice at a very young age is normally sufficient for PCR genotyping but not for a Southern Blot.

  6. In what way do thymus and adrenal gland weights reflect chronic stress in the host:

        1. Both weights decrease with stress

        2. Both weights increase with stress

        3. Thymus weight decreases but adrenal weight increases

        4. Thymus weight increases but adrenal weight decreases

ANSWERS:


  1. c. Postnatal days 5-7

  2. b. 1mm in diameter and 6mm long

  3. a. Anxiety

  4. d. Motor coordination and balance

  5. True

  6. c. Thymus weight decreases but adrenal weight increases



Kemppinen et al. Impact of aspen furniture and restricted feeding on activity, blood pressure, heart rate and faecal coritcosterone and immunoglobulin A excretion in rats (Rattus norvegicus) housed in individually ventilated cages, pp. 104-112
Domain 4: Animal Care, K1. Species-specific husbandry (e.g. nutrition, housing, exercise); K2. Environmental enrichment
Primary Species: Rat
SUMMARY: The objective of the study was to evaluate the impact of adding different items to cages on the rat’s activity, cardiovascular parameters and fecal stress indicators.  They compared three different aspen cage items:  a cross, diet board-a similar cross with drilled holes loaded with food pellets (no other food source was given), and a rectangular tube, as well as control group with no furniture.  Male rats of strains BN and F344 were housed in groups of three; one rat per group was implanted with a telemetric transponder to measure MAP and HR.  Rats were allowed 10 days recovery before experiment started.  Each group spent 14 days with cage furniture after this period rats were housed singly for 6 hours and all fecal pellets were collected and analyzed for corticosterone and IgA.  As a way of determining which of the statistically significant MAP and HR mean changes were biologically meaningful, the night-day differences of the controls on day 14 were used. F344 rats had consistently higher MAP and HR compared to BN rats, the dark activity levels of the F344 rats were also higher than those of BN.  F344 rats had the highest MAP in the in the tube cages, however it is possible the tube may have been too small to comfortably accommodate 3 large rats as this is only applicable to the F344 rats.  Both cross structures lowered MAP values for F344 rats, the plain was most effective.  BN had no biologically significant MAP or HR changes between furniture items.  No effect was seen on fecal corticosterone or IgA excretion compared to rats in control cages.  Establishment of general recommendations for cage furniture may be difficult because there is a clear genetic component involved, resulting in major differences between strains.  Cardiovascular parameters can be used to assess the welfare value of cage furniture, whereas changes in fecal corticosterone and fecal IgA excretion appear to be too small to be quantifiable. 
QUESTIONS:

  1. T/F European regulations mandate the use of nesting material or nest boxes in rodent cages.

  2. Which of the following strains of rat did cage items not significantly reduced heart rate? 

  1. F344

  2. SH

  3. BN

  4. SH

  5. C and D

  1. What dose injection of adrenocorticotropin hormone is required to detect a subsequent change in the fecal corticosteroid level in rats?

  1. 25 µg/kg

  2. 50 µg/kg

  3. 100 µg/kg

  1. T/F Rats blood pressure and heart rate are lower during the night than during the day.

ANSWERS:


  1. T

  2. E.  Sharp J, Azar T, Lawson D.  Effects of a cage enrichment program on heart rate, blood pressure, and activity of male Sprague-Dawley and spontaneously hypertensive rats monitored by radiotelemetry.  Contemp Top Lab Anim Sci 2005; 44:32-40

  3. C. Changes to the cage environment may not have stimulated the hypothalamic-pituitary-adrenal axis enough to produce detectable changes in fecal corticosteroid excretion.   Siswanto H, Hau J, Carlsson H-E, Goldkuhl R, Abelson KSP.  Corticosterone concentrations in blood and excretion in faeces after ATCH administration in male Sprague-Dawley rats.  In Vivo 2008; 22:435-440

  4. F. Rats are crepuscular and nocturnal and have increased HR and BP at night.



Allweiler et al. The use of propofol and sevoflurane for surgical anaesthesia in New Zealand White rabbits, pp. 113-118
Domain 2: Management of Pain and Distress; Task 2: Minimize or eliminate pain and/or distress. T2. Diagnose disease or condition as appropriate. Knowledge K5.
Primary Species: rabbits

 

SUMMARY: The purpose of this study was to evaluate anaesthesia for ovariohysterectomy in 34 New Zealand White rabbits, with induction of anaesthesia with propofol (16±5 mg/kg IV), and maintenance with sevoflurane (4.0±0.5%). The quality of anaesthetic induction and recovery from a surgical plane of anaesthesia was evaluated, and selected cardiopulmonary parameters (heart rate, respiratory rate, arterial blood pressure, haemoglobin oxygen saturation, end-tidal carbon dioxide concentration, end-tidal sevoflurane concentration and oesophageal temperature) effects were recorded. Time from induction of anaesthesia to tracheal extubation and sternal recumbency were recorded. The rabbits were part of a study aimed at developing a pain assessment scheme for rabbits. Propofol produced smooth induction of anaesthesia without production of apnea. Intubation was successfully performed in 33 rabbits. No ventilatory support was required during the anaesthetic period. No anaesthetic-related mortality occurred and all animals recovered uneventfully. Propofol administered previously to sevoflurane anaesthesia produced a good quality of surgical anaesthesia for ovariohysterectomy and stable cardiopulmonary conditions. This study indicates that induction of anaesthesia with propofol followed by maintenance with sevoflurane is a suitable technique for use in healthy rabbits undergoing ovariohysterectomy.


QUESTIONS:

    1. What minimum alveolar concentration (MAC) of sevoflurane has been reported in response to a noxious stimulus (tail clamp) in rabbits?

          1. 1.6%

          2. 2.4%

          3. 3.7%

          4. 4.0%

          5. All of them are correct

          6. All of them are false

    1. Responses of rabbis exposed to very low concentrations of volatile anaesthetics include:

  1. Hold their breath when exposed

  2. Bradycardia

  3. Hypercapnia

  4. Hypoxia

  5. All of them are correct

    1. The main advantage of propofol over other hypnotic agents in humans are:

  1. Its rapid metabolism

  2. No excitation during induction

  3. No excitation during recovery

  4. All of them are correct

  1. The lowest reported dose of propofol to produce apnea in rabbits is:

  1. 20 mg/kg

  2. 16 mg/kg

  3. 10 mg/kg

  4. 5 mg/kg

  1. Sevoflurane is a parenteral, flammable anaesthetic with pungent odor. True or False

  2. A study of anaesthetic mortality in veterinary practice highlighted the particularly high risk of death when anaesthetizing rabbits compared with dogs and cats, with the postoperative period being associated with the highest incidence of mortality. True or False.

ANSWERS:


1. c. (reported by Scheller et al., 1988. Can J Anaesth 35: 153-156.)

2. e. (reported by Hedenqvist et al., 2001. Lab Animal 35:172-179; Flecknell et al., 1996. Lab Animal 30:67-74; Flecknell et al., 1999. Lab Animal 33:41-46).

3. d. (reported by Biebuyk and Phil, 1989. Anesthesiology 71:261-277).

4. a. (reported by Glen JB, 1980. Br J Anesth 52:731-742).

5. False. Sevoflurane is a volatile, non-flammable anaesthetic with little or no pungent odor. Induction and recovery are rapid in comparison to any other volatile anaesthetics. (reported by Preckel and Bolten, 2005. Clin Anaesthesiol 19:331-348).

6. True. (reported by Brodbelt et al., 2008. Vet Anaesth and Analg 35:365-373).



Liljensten et al. Microdialysis as a method for biochemical and physiological studies of the porcine and human disc, pp. 119-123
Primary Species: Pig


SUMMARY: Actually, the physiological and the pathophysiological biochemistry of the disc are relatively unknown and not fully understood. No minimally invasive technique exists for the continuous evaluation of the biochemistry of animal and human intervertebral discs. In this paper, the microdialysis method is evaluated to see if it can could be used for research purposes as a new technique. It was performed in the lumbar discs of the pig to verify that a dialysate could be obtained by microdialysis, and in a human cadaver to establish de appropriate catheter dimensions for use in humans. Conventional SPF pigs (n=2) were used to evaluate cannulation and then the insertion of a 10 mm microdialysis membrane (n=6) guided with fluoroscopy to confirm catheter placement in the L3 and L4. The dialysate was recovered from the nucleus pulposus. Access to lumbar porcine disc was obtained with an 18 G 2 in. in a 35-45 º angle from the sagittal plane, and took less than 8 min. to perform. The flush rate was of 0.5 µL/min., and the dialysates could be recovered and glucose, lactate and pyruvate analyzed. In the human cadaver, the L4-L5 disc was accessed by a 19 G 3 in. needle with 35-40º angle. It was possible to apply a 30 mm membrane apart from the 10 mm one. The results obtained confirm that it is possible to obtain analyzable pyruvate; lactate and glucose metabolites from the nucleus pulposus in pig and human cadavers, and it encourage further in vivo studies using microdialysis technique on intervertebral discs.
QUESTIONS:

  1. (Find the False) With a 20 kDa membrane you can recover and analyze:

    1. Pyruvate

    2. Cytokines

    3. Lactate

    4. Glucose

  1. The dialysate is retrieved from the:

    1. Annulus fibrosus

    2. Nucleus pulposus

    3. Vertebral body

    4. Bone Marrow

  1. What is the gold thread used for?

    1. For making the membrane more flexible

    2. For confirming the position of the membrane in the nucleus pulposus

    3. For making the membrane more resistant

    4. None of these

  1. The composition of the perfusion fluid includes:

    1. Cl-

    2. Na+

    3. P+

    4. A and B

  1. True or False? The introduction of the needle and the dialysis membrane is guided with Magnetic Resonance Imaging.

  2. True or False? In the human cadaver, the cadaveric changes make the disc have a bigger size.

  3. True or False? The microdialysis is an easy and quick technique.

  4. True or False? In this study the microdialysis is used for the first time to monitor intervertebral discs in situ.

  5. True or False? The angle to introduce the needle into the nucleus pulposus is very different form the pig to the human.

  6. During the microdialysis, the dialysates could be recovered every:

    1. 2 min.

    2. 5 min.

    3. 15 min.

    4. 1 hr.

  1. True or False? Most disc herniation are localized in the lumbar region.

  2. True or False? Under post-mortem conditions it is not possible to obtain analyzable metabolites.

ANSWERS:


  1. B

  2. B

  3. B

  4. D

  5. False

  6. False

  7. True

  8. True

  9. False

  10. C

  11. True

  12. False



Murillo-Cuesta et al. Comparison of different aminoglycoside antibiotic treatments to refine ototoxicity studies in adult mice, pp. 124-131
Task 3: Research - Replacement, Reduction and Refinement Techniques

Primary Species: Mice


SUMMARY: Ototoxicity refers to the damage generated by a chemical substance in the organ responsible for hearing and balance. Aminoglycosides typically damage the outer hair cells (OHC) of the organ of Corti, beginning in the basal region of the cochlea and then spreading towards the middle and apical zones. Animal models used to study aminoglycoside toxicity have included the zebrafish, amphibians Rana catesbeiana and Xenopus laevis, chinchilla, rats and guinea pigs; in addition, the chinchilla has the same auditory frequency range as humans. The mouse is the preferred model to explore the basis of cochlear pathologies and new therapeutic interventions. In this study, four systemic treatments of aminoglycoside antibiotics gentamicin, neomycin, amikacin and kanamycin, were used to evaluate their effects on mortality, cochlear morphology and auditory brainstem responses. Two strains of mice were selected- HsdOla:MF1, an outbred strain and C57BL/6JOlaHsd, an inbred strain with the genetic predisposition to age-related hearing loss and high susceptibility to cochlear damage. This strain may carry a mutation in the Ahl gene which may contribute to otic insult susceptibility.  Hearing was evaluated by auditory brainstem responses (ABR) which is a non-invasive technique commonly used for hearing evaluation in mice. The results showed that gentamicin and amikacin caused high mortality without significantly changing the auditory threshold. Amikacin produced a tolerable rate of mortality but at doses that did not exhibit ototoxicity. IM injection of kanamycin in C57BL/6JOlaHsd mice induced significant dose-dependent bilateral hearing loss with a moderate rate of mortality and less discomfort than subcutaneous injection. 
QUESTIONS:

  1. Which mouse strain was predisposed to age-related hearing loss and high susceptibility of cochlear damage?

  2. Which route of administration of kanamycin induced significant dose-dependent bilateral hearing loss with a moderate rate of mortality and less discomfort?

ANSWERS:


  1. C57BL/6JOlaHsd

  2. Intramuscular injection


Takeo and Nakagata. Combination medium of cryoprotective agents containing L-glutamine and methyl-β-cyclodextrin in a preincubation medium yields a high fertilization rate for cryopreserved C57BL/6J mouse sperm, pp. 132-137
Primary Species: Mice
SUMMARY: The authors base the study in the fact that cryopreservation of mouse sperm provides a simple and economical means of storing the mice in mouse resource facilities as a vast number of genetically-engineered mice have been created in various laboratories worldwide recently. The protocol for sperm cryopreservation using 18% raffinose pentahydrate and 3% skim milk (R18S3) has been adopted in most laboratories. In general, relatively high fertilization rates can be obtained for frozen/thawed sperm in many inbred and F1 hybrid strains. However, the sperm of C57BL/6J mice shows an extremely low fertility rate after freezing and thawing (0–20%). In this study, they attempted to improve the low fertility of frozen/thawed C57BL/6J mouse sperm. The results showed that a combination of R18S3 containing L-glutamine and methyl-b-cyclodextrin (MBCD) in a preincubation medium dramatically increased the rate of fertilization (69.2+12.2%). Furthermore, the developmental potencies of two-cell embryos produced by frozen/thawed sperm to live young were normal (fresh: 46.0+8.2%, frozen/thawed: 51.5+11.1%). In summary, the authors conclude that their new method of sperm cryopreservation and in vitro fertilization using modified R18S3 with L-glutamine and MBCD in a preincubation medium yields a high fertilization rate for frozen/thawed C57BL/6J strain sperm. Furthermore, the new method provides a reliable archiving and reproducing system for genetically engineered mice using sperm cryopreservation.
QUESTIONS (Answer True or False):

  1. Generally speaking, frozen/thawed sperm in many inbred and F1 hybrid strains show relatively low fertilization rates in in vitro fertilization (IVF) including the sperm
    from C57BL/6J mice, which is the main strain used for the production of transgenic mice.

  2. Previously, the same authors have demonstrated that methylb-cyclodextrin (MBCD) dramatically improved the fertility of frozen/thawed sperm in C57BL/6J mice by stimulating the calcium influx to the spermatozoid flagellum.

  3. The authors have found that cryopreserved C57BL/6J mouse sperm frequently suffer from cryo-induced cellular injuries during freezing and thawing.

  4. There are many reports stating that L-glutamine acts as a cryoprotectant agent for sperm freezing and thawing in several mammalian species (human, bovine, etc…).

  5. Detailed mechanisms regarding the cryoprotective effect of L-glutamine on mouse sperm are very well known and there are several studies on the subject offering conclusive data.

ANSWERS:



  1. False: In general, frozen/thawed sperm in many inbred and F1 hybrid strains show relatively high fertilization rates in in vitro fertilization (IVF).However, the sperm
    from C57BL/6J mice, which is most used strain in the production of transgenic mice, show a low level of fertility after freezing and thawing (0–20%).

  2. False: It has been demonstrated that methylb-cyclodextrin (MBCD) dramatically improved the fertility of frozen/thawed sperm in C57BL/6J mice by stimulating the cholesterol efflux from the plasma membrane.

  3. True

  4. True

  5. False: Detailed mechanisms regarding the cryoprotective effect of L-glutamine on mouse sperm are not fully clear yet, so further studies will be required in the future.



Gokbulut et al. Plasma dispositions of ivermectin, doramectin and moxidectin following subcutaneous administration in rabbits, pp. 138-142
Domain 3: Research

 

Primary Species: Rabbit (Oryctolagus cuniculus)



 

SUMMARY: This study compared the plasma dispositions of ivermectin (IVM), doramectin (DRM) and moxidectin (MXD) following subcutaneous injection in rabbits.  15 six-month-old NZW rabbits (9 males and 6 females) with body weights ranging from 2.5-3.4kg were injected subcutaneously in the lateral midline of the back with a single dose of 0.3mg/kg IVM, DRM, or MXD.  Blood samples were collected at various times from 1 hour to 40 days post injection.  HPLC using fluorescence was used to analyze the samples.  MDX was absorbed faster and reached the peak plasma concentration (Cmax) significantly earlier than IVM or DRM.  There was no significant difference in Cmax between the three groups.

The persistence of DRM and MXD were significantly longer than IVM (30 days vs. 20 days, respectively).  Parameters did not vary between genders.  The authors speculate differences may have been due to formulations: MXD is in aqueous solution, while IVM is in glycol and DRM is in oil.


QUESTIONS:

    1. T or F?  MXD is 100 times more lipophilic than IVM.

    2. Avermectins and milbemycins are endectocides.  What would you use them to treat?

    1. Bacteria

    2. Viruses

    3. Nematodes and arthropods

    4. Protozoa 

 

ANSWERS:


      1. T.  MXD is also higher in water solubility.

      2. c

 
Khalifeh et al. Investigations of the role of tumor necrosis factor-α, interleukin-1β, interleukin-10, nitric oxide and rheumatoid factor-immunoglobulin M in a rat model of arthritis, pp. 143-149
Primary Species: Rat


SUMMARY: Collagen-induced arthritis (CIA) in rats is a widely used preclinical animal model of rheumatoid arthritis (RA). However, CIA development in SD rats is less severe in terms of inflammatory response compared with other strains. To solve this, a modified CIA model using N-acetylmuramyl dipeptide (MDP) called MCIA was developed for use in the less sensitive SD rat strains. This work was conducted to better understand the immunopathological role and contributions of the pro-inflammatory T-helper type 1 (Th-1) cytokines and inflammatory mediators, interleukin-1 (IL-1 β), tumour necrosis factor-alpha (TNF-α) and nitric oxide (NO); the anti-inflammatory T-helper type 2 (Th-2) cytokine, IL-10 and autoantibodies such as rheumatoid factor (RF)-immunoglobulin M (IgM) in this newly developed RA model. TNF-α, NO and RF-IgM levels were significantly increased, while IL-1β levels were not affected in this MCIA rat model. The levels of IL-10 were lower than the baseline when compared with controls.
In conclusion:

(1) The immunological features represented in the MCIA rat model favors the Th-1 cytokine profile over Th-2

(2) RF-IgM can be used as a diagnostic test in preclinical RA models.
QUESTIONS:

1. True or False? Serum IgM level is an important biomarker in collagen-induced arthritis rat model.



2. True or False? There are only limited preclinical models of rheumatoid arthritis.

3. True or False? IL-10 has potent anti-inflammatory functions and is reported to reduce joint swelling, cellular infiltration, pro-inflammatory cytokine production and cartilage degradation in collagen-induced arthritis in rats.

4. True or False? RF-IgM cannot be used as a diagnostic test in preclinical rheumatoid arthritis models.
ANSWERS:

1. True.

2. False. There are several.

3. True.



4. False. RF-IgM can

Honjo et al. Kinetics of α2-macroglobulin and α1-acid glycoprotein in rats subjected to repeated acute inflammatory stimulation, pp. 150-154
SUMMARY: The kinetics of α2-macroglobulin (α2M) and α1-acid glycoprotein (AAG) in rats repeatedly stimulated with intramuscular injections of turpentine oil at doses 0.05 and 0.4 mL/rat were investigated. Mean serum levels of α2M peaked at 48 h after the first turpentine oil injection, reaching 1.74 and 2.36 mg/mL at 0.05 and 0.4 mL/rat, respectively. AAG peaks were also observed at 48 h after injection, and the mean values were 2.02 and 2.53 mg/mL, respectively. These peak values of α2M and AAG differed significantly between the 0.05 and 0.4 mL/rat injection groups. Mean serum levels of interleukin-6 (IL-6) at 0.05 mL/rat were 52.61 pg/mL at 12 h, 48.86 pg/mL at 36 h and 81.93 pg/mL at 84 h after the first injection. Mean IL-6 serum levels at 0.4 mL/rat were 215.24 pg/mL at 12 h, 56.33 pg/mL at 36 h and 39.25 pg/mL at 84 h after the first injection. Mean serum levels of cytokine induced chemoattractant-1 (CINC-1) at a dose of 0.05 mL/rat were 5.70 ng/mL at 12 h, 5.58 ng/mL at 36 h and 4.58 ng/mL at 84 h after the first injection. Mean serum levels of CINC-1 after injection at 0.4 mL/rat were 11.57 ng/mL at 12 h, 4.68 ng/mL at 36 h and 4.42 ng/mL at 84 h. Serum levels of IL-6 differed significantly at 12, 24, 72 and 84 h, while those of CINC-1 differed significantly at 12, 24, 48 and 96 h between the 0.05 and 0.4 mL/rat injection groups. Differences in peak serum levels in the 0.05 and 0.4 mL/rat groups were attributed to differences in the production of IL-6 and CINC-1, which are thought to contribute to α2M and AAG production.

 

QUESTIONS: 



  1. True or false: serum levels of acute-phase proteins decrease in response to acute inflammation, infectious diseases, surgical trauma or pregnancy.

  2. Which proteins are used as typical acute-phase proteins in rats?

  3. True or False: the changes in serum levels of α2M and AAG after repeated acute inflammatory stimulation at doses of 0.05 and 0.4 mL/rat showed a similar pattern to single stimulation.

  4. True or false: differences in peak serum levels in the 0.05 and 0.4 mL/rat groups were attributed to differences in the production of IL-6 and CINC-1, which are thought to contribute to α2M and AAG production.

 

ANSWERS: 



  1. False: these levels increase in these situations.

  2. α2-macroglobulin (α2M) and α1-acid glycoprotein (AAG).

  3. True.

  4. True.

 
Ozpinar et al. Thyroidal radioactive iodide uptake in the lactating rhesus monkey, pp. 155-158
Domain 1, Task 3

Primary Species: Macaques (Macaca spp.)

 

SUMMARY: Radioactive iodide uptake (RAIU) assesses gland activity and hormone synthesis integrity in the thyroid. It is used to evaluate thyroid function and diagnosis thyroid disease in humans. Percentage of iodide uptake will vary with iodine status. RAIU is generally performed 24 hrs after single dose oral administration of 123I for better diagnostic accuracy. The iodide ion is readily absorbed from the stomach. The RAIU test measures radioactive iodine accumulation in the thyroid at different time intervals using a gamma scintillation counter. 123I or 131I are commonly used. 123I is the radionucleotide of choice since it is associated with less radiation dose than 131I; it is especially attractive for use in infants and pregnant or lactating women. 123I has high specific activity and is virtually carrier free. RAIU is the most widely used test in human clinical nuclear medicine labs. 123 I scan is more reliable for evaluation of thyroid anatomy and function than 99mTcO4-4.  Rhesus monkeys have been used as a model for the study of pregnancy, postnatal growth and lactation. They are considered the best model for thyroid function assessment especially with the similarities between monkeys and human infants.  This study sought to validate the RAIU test for lactating rhesus monkeys in order to further human model studies, especially those involving pregnant or lactating females and infants for which there is a dearth of information. RAIU was performed in 4 lactating rhesus macaques. Thyroidal RAIU varied significantly among the animals even though baseline thyroid function tests, T3, T4 & TSH hormone levels, were normal. The authors believe the RAIU test may allow a rational clinical approach to thyroid function testing in lactating rhesus, however additional studies are needed to develop appropriate age/gender “normals” and determine what results to expect from normal versus clinically abnormal animals.
QUESTIONS: 


    1. Radioactive iodide uptake (RAIU) tests:

    1. Assess integrity of hormone biosynthesis in the thyroid gland

    2. Are commonly used to evaluate thyroid function and diagnose thyroid disease

    3. Assess the level of thyroid gland activity

    4. All of the above

    1. 123I is generally administered:

          1. Orally

          2. Intravenously

          3. Topically

          4. None of the above

    1. T/F:  The RAIU test uses a gamma scintillation counter to measure radioactive iodine accumulation in the thyroid gland over time.

 

ANSWERS: 



      1. d

      2. a

      3. T


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