The PTPN13 Y2081D (T>G) (rs989902) polymorphism is associated with an increased risk of sporadic colorectal cancer.

AIM: Colorectal cancer (CRC) is one of the most common cancers worldwide and, although the majority of cases are sporadic, its development and progression depends on a range of factors: environmental, genetic and epigenetic. A variety of genetic pathways have been described as being crucial in CRC, including protein tyrosine phosphatases (PTPs). PTPN13 (also called FAP-1) is a non-receptor PTP and interacts with a number of important components of growth and apoptosis pathways. It is also involved in the inhibition of Fas-induced apoptosis. METHOD: The single nucleotide polymorphism genotype at Y2081D (T>G) (rs989902) of PTPN13 exon 39 was determined in DNA extracted from blood samples from 174 sporadic CRC patients and 176 healthy individuals. Also, a meta-analysis was performed based on three articles accessed via the PubMed and ResearchGate databases. RESULTS: The risk of CRC was 2.087 times greater for patients with the GG genotype than for those with the TT genotype (P = 0.0475). In the meta-analysis, a significantly increased risk of cancer associated with the G allele was observed in the squamous cell carcinoma of the head and neck subgroup (TT vs GG+GT, OR 1.23, 95% CI [1.02, 1.47], P = 0.0258), and a significantly decreased risk in the breast cancer subgroup (TT vs GG+GT, OR 0.63, 95% CI [0.41, 0.96], P = 0.0334) and in the CRC subgroup (GT+TT vs GG, OR 0.51, 95% CI [0.41, 0.95], P = 0.0333). CONCLUSION: PTPN13 rs989902 is significantly associated with the risk of CRC in the Polish population. Given that this report provides the first evidence of an association of PTPN13 rs989902 with the risk of CRC in a Caucasian population, further large scale studies are necessary to confirm this finding.

Effects of temporary access to environmental enrichment on measures of laboratory mouse welfare.

Laboratory mice are typically housed in "shoebox" cages with limited opportunities to engage in natural behaviour. Temporary access to environments with increased space and complexity (playpens) may improve mouse welfare. Previous work by our group has shown that mice are motivated to access and use these environments, but it is unknown how other aspects of welfare are impacted. Female C57BL/6J, BALB/cJ, and DBA/2J mice (n = 21; 7 mice per strain) were housed in mixed-strain trios and given temporary access to a large playpen with their cage mates three times per week. Control mice (n = 21; 7 mice per strain) remained in their home cages. Home cage behaviour (development of stereotypic behaviour over time, aggression following cage-changing) and anxiety tests were used to assess how playpen access impacted welfare. Contrary to our predictions, we found increased time spent performing stereotypies in playpen mice; this difference may be related to negative emotional states, increased motivation to escape the home cage, or active coping strategies. Playpen access resulted in strain-dependent improvements in aggression and some measures of anxiety. Aggression was lower for C57BL/6J mice in the playpen treatment following cage changing than it was for C57BL/6J control mice, while playpen mice, and particularly the C57BL/6J strain, spent more time in the center of the open field test and produced fewer fecal boli during anxiety testing, supporting other research showing that strain differences play an important role in behaviour and stress resiliency.

Rat aversion to isoflurane versus carbon dioxide.

Some experts suggest that sedation of laboratory rodents with isoflurane before euthanasia with carbon dioxide (CO(2)) is a humane alternative to euthanasia with CO(2) alone, but little research has compared aversion with these agents. Albino rats were tested in a light-dark box where they had the choice between remaining in a dark compartment filling with isoflurane or CO(2), or escaping to a lit compartment. Experiment 1 validated the procedure by confirming that rats responded to agent and light intensity. In experiment 2, 9/16 and 0/16 rats remained in the dark compartment until recumbent when initially exposed to isoflurane and CO(2), respectively. In experiment 3, more rats remained in the dark compartment until recumbent during initial (10/16) versus re-exposure (1/16) to isoflurane. These results indicate that initial exposure to CO(2) is more aversive than isoflurane, and that re-exposure to isoflurane is more aversive than initial exposure. We conclude that sedation with isoflurane is a refinement over euthanasia with CO(2) alone for rats that have not been previously exposed to inhalant anaesthetics.

A Good Life for Laboratory Rodents?

Most would agree that animals in research should be spared "unnecessary" harm, pain, or distress, and there is also growing interest in providing animals with some form of environmental enrichment. But is this the standard of care that we should aspire to? We argue that we need to work towards a higher standard-specifically, that providing research animals with a "good life" should be a prerequisite for their use. The aims of this paper are to illustrate our vision of a "good life" for laboratory rats and mice and to provide a roadmap for achieving this vision. We recognize that several research procedures are clearly incompatible with a good life but describe here what we consider to be the minimum day-to-day living conditions to be met when using rodents in research. A good life requires that animals can express a rich behavioral repertoire, use their abilities, and fulfill their potential through active engagement with their environment. In the first section, we describe how animals could be housed for these requirements to be fulfilled, from simple modifications to standard housing through to better cage designs and free-ranging options. In the second section, we review the types of interactions with laboratory rodents that are compatible with a good life. In the third section, we address the potential for the animals to have a life outside of research, including the use of pets in clinical trials (the animal-as-patient model) and the adoption of research animals to new homes when they are no longer needed in research. We conclude with a few suggestions for achieving our vision.

Using approach latency and anticipatory behaviour to assess whether voluntary playpen access is rewarding to laboratory mice.

Laboratory mice are typically housed in "shoebox" cages that limit the expression of natural behaviours. Temporary access to more complex environments (playpens) may improve their welfare. We aimed to assess if access to playpens is rewarding for conventionally-housed mice and to document mouse behaviour during playpen access. Female C57BL/6J, BALB/cJ, and DBA/2J mice were provided temporary access to a large enriched playpen three times per week; control mice remained in their home cages. We measured latency to enter playpens and anticipatory behaviour to determine if access was rewarding, and recorded mouse behaviour during playpen sessions. Over time, playpen mice entered the playpen more quickly; latency declined from 168 ± 22 to 13 ± 2 s over the 14-d trial. As expected, playpen mice showed an increase in anticipatory behaviour before playpen access (mean ± SE = 19.7 ± 2.6 behavioural transitions), while control mice showed no change in anticipatory behaviour relative to baseline values (2.4 ± 1.6 transitions). Mice in the playpen performed more ambulatory behaviours than control mice who remained in home cages (21.5 ± 0.7 vs 6.9 ± 1.1 observations of 25 total observations). We conclude that conventionally-housed mice find voluntary playpen access rewarding, and suggest this as a useful option for providing laboratory mice with access to more complex environments.

Standard laboratory housing for mice restricts their ability to segregate space into clean and dirty areas.

Laboratory mice (Mus musculus) are typically housed in simple cages consisting of one open space. These standard cages may thwart mouse ability to segregate resting areas from areas where they eliminate, a behaviour that is prevalent across the animal kingdom. No scientific work has directly tested whether mice engage in such segregation behaviour, or whether the ability to do so may have welfare consequences. Here we show that mice, whether housed in standard cages or a complex housing system consisting of three interconnected standard cages, kept nesting and elimination sites highly segregated, with nest and urine co-occurring in the same location only 2% of the time. However, mice in the complex system established these clean and dirty sites in separate cages instead of separate locations within one cage, and carried bedding materials (cellulose pellets) from their nesting cages to their latrine cage. Moreover, mice in the complex system displayed more behaviours associated with positive welfare and were less disturbed by weekly husbandry procedures. We conclude that mice find waste products aversive, and that housing mice in a way that facilitates spatial segregation provides a simple way of allowing the expression of natural behaviours and improving welfare.

Correction to 'The importance of burrowing, climbing and standing upright for laboratory rats'.

[This corrects the article DOI: 10.1098/rsos.160136.].

Myelodysplastic syndromes according to FAB and WHO classification. Single center experience.

The results of clinical and laboratory observations of 119 MDS patients divided acc. to FAB, and - after excluding RAEB-t and CMML groups -- of 95 patients divided accordingly to WHO classification are presented. The diagnosis of MDS was based on medical interview, physical examination, blood biochemistry, peripheral blood (PB) and bone marrow (BM) cytomorphology and cytochemistry, trephine biopsy and cytogenetic examination. All hematologic examinations were done according to routine methods. Cytogenetic analyses were carried out on BM cells from 24-48 h cultures in standard conditions. At least 15-20 GTG-banded metaphases were analyzed in every patient. The survival time (ST) of patients differed significantly between the FAB or WHO groups, with p=0.0004 for FAB and p=0.02 for WHO. The progression to AML was more common in less favorable groups, with p=0.0001 for FAB and p=0.00016 for WHO. The distribution of IPSS prognostic index among the groups showed statistically significant difference (p=0.0004 for FAB, and p=0.0001 for WHO), whereas the distribution of karyotypic abnormalities did not. However, in univariate analysis statistically significant influence on ST showed, beside the both classification systems: cytogenetics, the presence of blasts in PB, age and IPSS index. In multivariate analysis the sole independent prognostic factors were: PB blasts and cytogenetics. The authors conclude that the WHO classification offers a good prognostic tool for MDS patients. However, the karyotype and the presence of blasts in PB should always be taken into account.

The importance of burrowing, climbing and standing upright for laboratory rats.

Standard laboratory cages prevent rats (Rattus norvegicus) from performing many behaviours that they perform in the wild, but little is known about how this may affect their welfare. The aims of this study were (i) to record the propensity to burrow, climb and stand upright in 3-, 8- and 13-month old laboratory rats housed in semi-naturalistic environments and (ii) to compare the frequency of lateral stretching in semi-naturalistic versus standard-housed rats; we predicted standard-housed rats would perform more lateral stretches to compensate for the inability to stretch upright. Rats' propensity to burrow remained constant as they aged (approx. 30 bouts per day totalling 20-30 min), suggesting burrowing is important to rats. Climbing decreased from 76 to 7 bouts per day at 3 versus 13 months, probably because of declining physical ability. Upright standing decreased from 178 to 73 bouts per day, but continued to be frequently expressed even in older rats. Standard-housed rats stretched much more frequently than semi-naturalistic-housed rats (53 versus 6 bouts per day at 13 months), perhaps in compensation for inability to stretch upright and to relieve stiffness caused by low mobility associated with standard housing. These findings suggest that standard laboratory cages interfere with important natural behaviours, which is likely to compromise rat welfare.

Differences in Anticipatory Behaviour between Rats (Rattus norvegicus) Housed in Standard versus Semi-Naturalistic Laboratory Environments.

Laboratory rats are usually kept in relatively small cages, but research has shown that they prefer larger and more complex environments. The physiological, neurological and health effects of standard laboratory housing are well established, but fewer studies have addressed the sustained emotional impact of a standard cage environment. One method of assessing affective states in animals is to look at the animals' anticipatory behaviour between the presentation of a cue signalling the arrival of a reward and the arrival of that reward. The primary aim of this study was to use anticipatory behaviour to assess the affective state experienced by female rats a) reared and housed long-term in a standard laboratory cage versus a semi-naturalistic environment, and b) before and after treatment with an antidepressant or an anxiolytic. A secondary aim was to add to the literature on anticipatory behaviour by describing and comparing the frequency and duration of individual elements of anticipatory behaviour displayed by rats reared in these two systems. In all experiments, total behavioural frequency was higher in standard-housed rats compared to rats from the semi-naturalistic condition, suggesting that standard-housed rats were more sensitive to rewards and experiencing poorer welfare than rats reared in the semi-naturalistic environment. What rats did in anticipation of the reward also differed between housing treatments, with standard-housed rats mostly rearing and rats from the semi-naturalistic condition mostly sitting facing the direction of the upcoming treat. Drug interventions had no effect on the quantity or form of anticipatory behaviour, suggesting that the poorer welfare experienced by standard-housed rats was not analogous to depression or anxiety, or alternatively that the drug interventions were ineffective. This study adds to mounting evidence that standard laboratory housing for rats compromises rat welfare, and provides further scientific support for recommendations that current minimum standards be raised.

A Good Death? Report of the Second Newcastle Meeting on Laboratory Animal Euthanasia.

Millions of laboratory animals are killed each year worldwide. There is an ethical, and in many countries also a legal, imperative to ensure those deaths cause minimal suffering. However, there is a lack of consensus regarding what methods of killing are humane for many species and stages of development. In 2013, an international group of researchers and stakeholders met at Newcastle University, United Kingdom to discuss the latest research and which methods could currently be considered most humane for the most commonly used laboratory species (mice, rats and zebrafish). They also discussed factors to consider when making decisions about appropriate techniques for particular species and projects, and priorities for further research. This report summarises the research findings and discussions, with recommendations to help inform good practice for humane killing.

DNA methylation analysis of a de novo balanced X;13 translocation in a girl with abnormal phenotype: evidence for functional duplication of the whole short arm of the X chromosome.

We report on a 13-month-old girl showing dysmorphic features and a delay in psychomotor development. She was diagnosed with a balanced de novo translocation 46,X,t(X;13)(p11.2;p13) and non-random inactivation of the X chromosome. FISH analysis, employing the X chromosome centromere and XIST-region-specific probes, showed that the XIST locus was not involved in the translocation. Selective inactivation of paternal X, which was involved in translocation, was revealed by the HUMARA assay. The pattern of methylation of 5 genes located within Xp, which are normally silenced on an inactive X chromosome, corresponded to an active (unmethylated) X chromosome. These results revealed that in our proband the X chromosome involved in translocation (Xt) was preferentially inactivated. However, genes located on the translocated Xp did not include XIST. This resulted in functional Xp disomy, which most probably accounts for the abnormal phenotype in our patient.

[Cytogenetic changes in Ph(+) chronic granulocytic leukemia under the influence of interferon alpha (IFN-alpha) therapy]. / Zmiany cytogenetyczne w Ph-dodatniej [Ph(+)] przewleklej bialaczce szpikowej w czasie leczenia interferonem alfa (IFN-alpha).

Chromosomal changes during therapy with IFN-alpha were analysed in 21 patients suffering of chronic granulocytic leukemia. Complete or major cytogenetic response (CgR) was obtained in 4 patients, minor in 5 minimal in 7, and no response in 5 patients. Patients showing a low Sokal index more often disclosed CgR. In 6 persons additional chromosomal aberrations were present at diagnosis or during the disease course. They disappeared on IFN-alpha therapy. This fact may be in favour of the influence of IFN-alpha not only on disappearance of Ph chromosome, but also of secondary aberrations, some of them indicating the possibility of an acceleration of the disease. No relation of the CgR, as well as Sokal index value to the survival time in presented small cohort could be observed, which may depend on short observation time. CgR did not depend on a type of fusion of BCR/ABL gene. However, survival time was longer in patients with b3a2 fusion.