Assessment of post-laparotomy pain in laboratory mice by telemetric recording of heart rate and heart rate variability

BACKGROUND: Pain of mild to moderate grade is difficult to detect in laboratory mice because mice are prey animals that attempt to elude predators or man by hiding signs of weakness, injury or pain. In this study, we investigated the use of telemetry to identify indicators of mild-to-moderate post-laparotomy pain. RESULTS: Adult mice were subjected to laparotomy, either combined with pain treatment (carprofen or flunixin, 5mg/kg s/c bid, for 1 day) or without pain relief. Controls received anesthesia and analgesics or vehicle only. Telemetrically measured locomotor activity was undisturbed in all animals, thus confirming that any pain experienced was of the intended mild level. No symptoms of pain were registered in any of the groups by scoring the animals' outer appearance or spontaneous and provoked behavior. In contrast, the group receiving no analgesic treatment after laparotomy demonstrated significant changes in telemetry electrocardiogram recordings: increased heart rate and decreased heart rate variability parameters pointed to sympathetic activation and pain lasting for 24 hours. In addition, core body temperature was elevated. Body weight and food intake were reduced for 3 and 2 days, respectively. Moreover, unstructured cage territory and destroyed nests appeared for 1-2 days in an increased number of animals in this group only. In controls these parameters were not affected. CONCLUSIONS: In conclusion, real-time telemetric recordings of heart rate and heart rate variability were indicative of mild-to-moderate post-laparotomy pain and could define its duration in our mouse model. This level of pain cannot easily be detected by direct observation

TGFbeta receptor II gene deletion in leucocytes prevents cerebral vasculitis in bacterial meningitis

In bacterial meningitis, chemokines lead to recruitment of polymorphonuclear leucocytes (PMN) into the CNS. At the site of infection in the subarachnoid space, PMN release reactive oxygen species, reactive nitrogen intermediates (RNI) and interleukin-1beta (IL-1beta). Although these immune factors assist in clearance of bacteria, they also result in neuronal injury associated with meningitis. Transforming growth factor beta (TGFbeta) is a potent deactivator of PMN and macrophages since TGFbeta suppresses the production of ROI, RNI and IL-1. Here, we report that the deletion of the TGFbeta receptor II gene in PMN enhances PMN recruitment into the CNS of mice with Streptococcus pneumoniae meningitis. This was associated with more efficient clearance of bacteria, and almost complete prevention of intracerebral necrotizing vasculitis. Differences in PMN in the CNS of infected control mice and mice lacking TGFbeta receptor II were not explained by altered expression of chemokines acting on PMN. Instead, TGFbeta was found to impair the expression of L (leucocyte)-selectin on PMN from control mice but not from mice lacking TGFbeta receptor II. L-selectin is known to be essential for PMN recruitment in bacterial meningitis. We conclude that defective TGFbeta signalling in PMN is beneficial in bacterial meningitis by ameliorating migration of PMN and bacterial clearance

Assessment of post-laparotomy pain in laboratory mice by telemetric recording of heart rate and heart rate variability-1

<p><b>Copyright information:</b></p><p>Taken from "Assessment of post-laparotomy pain in laboratory mice by telemetric recording of heart rate and heart rate variability"</p><p>http://www.biomedcentral.com/1746-6148/3/16</p><p>BMC Veterinary Research 2007;3():16-16.</p><p>Published online 2 Aug 2007</p><p>PMCID:PMC1965463.</p><p></p>eviation of interbeat interval [SDNN, milliseconds (ms)] relative to baseline (i.e., normal values taken the day before the experiment) are plotted over time. Symbols indicate 12-hour means (bars indicate ± SEM). Corresponding control experiments in which animals received anesthesia and injections only are depicted as black horizontal lines with grey columns representing ± SEM. Asterisks indicate statistical significance (= 8, paired Student's test with Bonferroni correction) at ≤ 0.008. Note increased heart rate values with decreased heart rate variability parameters [IBI, SDNN] during the first light phase (12–24 h) in operated animals without pain treatment

Assessment of post-laparotomy pain in laboratory mice by telemetric recording of heart rate and heart rate variability-3

<p><b>Copyright information:</b></p><p>Taken from "Assessment of post-laparotomy pain in laboratory mice by telemetric recording of heart rate and heart rate variability"</p><p>http://www.biomedcentral.com/1746-6148/3/16</p><p>BMC Veterinary Research 2007;3():16-16.</p><p>Published online 2 Aug 2007</p><p>PMCID:PMC1965463.</p><p></p>t the mean values from 8 animals, with bars indicating ± SEM. Corresponding control experiments in which animals received anesthesia and injections only are depicted as black horizontal lines with grey columns representing ± SEM. Asterisks indicate statistical significance (paired Student's test with Bonferroni correction) at ≤ 0.016. Note the reduction in body weight for three days and the decrease in food consumption for two days after laparotomy without pain treatment

Assessment of post-laparotomy pain in laboratory mice by telemetric recording of heart rate and heart rate variability-4

<p><b>Copyright information:</b></p><p>Taken from "Assessment of post-laparotomy pain in laboratory mice by telemetric recording of heart rate and heart rate variability"</p><p>http://www.biomedcentral.com/1746-6148/3/16</p><p>BMC Veterinary Research 2007;3():16-16.</p><p>Published online 2 Aug 2007</p><p>PMCID:PMC1965463.</p><p></p>ddings surface (circles). The lower row illustrates Score 1 with an unstructured cage area and two nest-like resting places (arrows)

Expression profiling in transgenic FVB/N embryonic stem cells overexpressing STAT3-2

the morula and of the blastocyst. Expression of Hexokinase II, Lefty2 and PP1s15B was restricted to the cells of the inner part of the morula and to the ICM in the blastocyst. Sense probes in the same concentration of the antisense probes were used as negative controls for the hybridization. In situ hybridization with a Pramel7 antisense riboprobe of preimplantation embryos (a: two-cell embryo, b: four-cell embryo, c: eight-cell embryo; d, e: compacted morula, f, g: blastocyst, h: negative control with sense riboprobe). Magnification 40×.<p><b>Copyright information:</b></p><p>Taken from "Expression profiling in transgenic FVB/N embryonic stem cells overexpressing STAT3"</p><p>http://www.biomedcentral.com/1471-213X/8/57</p><p>BMC Developmental Biology 2008;8():57-57.</p><p>Published online 23 May 2008</p><p>PMCID:PMC2409313.</p><p></p

Expression profiling in transgenic FVB/N embryonic stem cells overexpressing STAT3-3

Formed after cultivation of the cells for 8 days without adding LIF to the medium. E14 ES cells differentiated under these conditions and lost the expression of OCT-3/4 and SSEA-1 and alkaline phosphatase. Both Pem/Rhox5 and Pramel7 overexpressing cells maintained the expression of the pluripotency markers. Nanog overexpressing cells, used as a control for the experiment, as expected maintained their pluripotency. Scale bar in both large and small panels: 100 μm.<p><b>Copyright information:</b></p><p>Taken from "Expression profiling in transgenic FVB/N embryonic stem cells overexpressing STAT3"</p><p>http://www.biomedcentral.com/1471-213X/8/57</p><p>BMC Developmental Biology 2008;8():57-57.</p><p>Published online 23 May 2008</p><p>PMCID:PMC2409313.</p><p></p

Expression profiling in transgenic FVB/N embryonic stem cells overexpressing STAT3-1

-1 (A) and alkaline phosphatase (B). The expression of all markers was restricted to ES cells. Mouse fibroblast used as feeder cells are negative for OCT-3/4, SSEA-1 and alkaline phosphatase. Scale bar in both large and small panels: 100 μm.<p><b>Copyright information:</b></p><p>Taken from "Expression profiling in transgenic FVB/N embryonic stem cells overexpressing STAT3"</p><p>http://www.biomedcentral.com/1471-213X/8/57</p><p>BMC Developmental Biology 2008;8():57-57.</p><p>Published online 23 May 2008</p><p>PMCID:PMC2409313.</p><p></p

Expression profiling in transgenic FVB/N embryonic stem cells overexpressing STAT3-0

TAT3. After 24 hours of LIF and OHT deprivation cells were cultivated till their homogenization in presence of either LIF or OHT. During LIF or OHT deprivation no changes in the protein expression levels could be detected, but after 24 hrs the Tyr705 residue of both transgenic and WT STAT3 was completely dephosphorylated. 10 minutes after addition of LIF the tyr705 residue of both STAT3 was phosphorylated whereas after addition of OHT complete phosphorylation was obtained only after 6 hrs. Dephosphorylation of Tyr705 was analyzed by eliminating LIF or OHT from respectively WT or 743 cells. Kinetics for the dephosphorylation were slower then for the phosphorylation, only after 48 hrs dephosphorylation of WT Tyr705 was complete whereas complete dephosphorylation of STAT3-MER occurred only after 72 hrs. Nanog and STAT3 expression levels were tested by RTQ-PCR. Values are normalized with β-actin and the values indicate fold changes compared to the WT. Both WT and 743 ES cell lines expressed Nanog and the transgenic 743 ES cells have an increased Nanog expression compared to WT. After injection of 743 in C57BL/6 host blastocysts a 50–60% chimera was generated. Littermates from the crossing of the chimera with a WT FVB/N female generated white littermates, 50% of which were hemizygous for the transgene, indicating germline competence of the 743 cell line.<p><b>Copyright information:</b></p><p>Taken from "Expression profiling in transgenic FVB/N embryonic stem cells overexpressing STAT3"</p><p>http://www.biomedcentral.com/1471-213X/8/57</p><p>BMC Developmental Biology 2008;8():57-57.</p><p>Published online 23 May 2008</p><p>PMCID:PMC2409313.</p><p></p