Transitioning from crates to free farrowing: A roadmap to navigate key decisions.

There are animal welfare concerns about the continued use of permanent crating systems for farrowing and lactating sows, which is the most prevalent maternity system in global pig production. Greater societal attention in recent years has culminated in changes (or proposed changes) to regulations as well as market-driven initiatives to move away from crated systems. Transitioning from farrowing crates to systems that allow the sow greater freedom of movement and behavioral expression requires a number of key decisions, with various trade-offs apparent when trying to balance the needs of different stakeholders. This review discusses these decisions based on common questions asked by farmers, policy makers and other stakeholders when deciding on a new system to build/approve. Based on the latest scientific evidence and practical insight, decisions such as: whether to retrofit an existing barn or build a new one, what spatial dimensions are necessary per sow place, whether to adopt free farrowing or temporary crating, how to provide substrate/enrichment and be hygienic and environmentally friendly, and how to optimize the human inputs and transition between systems are considered. The aim of this paper is to provide a roadmap for those interested in uptake of higher welfare systems and practices, as well as to highlight areas requiring further optimization and research.

Review of Temporary Crating of Farrowing and Lactating Sows.

Temporary crating (TC) provides lactating sows with the opportunity to move more freely after crate opening a few days after parturition. The aim of this paper was to evaluate whether TC gives overall welfare improvement when compared to permanent crating or free farrowing. This review shows that when pens with TC allow the sows to turn during the majority of time in the farrowing unit, it is the pen design and period of confinement in a crate within it that influence the extent to which different functional and motivated behaviors can be fulfilled. This review also indicates that there are at least short-term benefits to sows when confinement is reduced, as shown by reported increases in motivated behaviors such as exploration and interactions with piglets when not permanently crated. It remains unclear whether there are any longer-term beneficial effects (until or beyond weaning) due to the paucity of studies. Furthermore, it is uncertain whether the observed short-term benefits translate to other welfare indicators. Research findings indicate no reduction in the frequency of stereotypies or body lesions and do not provide a clear answer regarding sow stress response when released from confinement. Compared to free farrowing, TC appears beneficial for reducing piglet mortality. The impact of the time of onset of TC on the farrowing process and piglet mortality have been inconsistent. While confinement before farrowing prevents nest building behavior, consequences of this for sow physiology have been ambiguous. Confining the sow briefly after farrowing may be the best compromise, allowing the sow to perform motivated nest-building behavior, but the risks of crushing during the unconfined farrowing period may increase. Subsequent crate reopening seems to increase piglet mortality but only if done earlier than 3-5 days after farrowing. The review also provides methodological considerations, a proposal for consistent and accurate terminology when describing systems and highlights gaps of knowledge. In conclusion, TC is a step forward to better pig welfare compared to the farrowing crate, as it allows some freedom of movement for sows without impairing piglet welfare. However, more comprehensive research is needed to draw sound conclusions as to whether TC is a viable transition from permanent crating to free farrowing.

Molecularly matched therapy in the context of sensitivity, resistance, and safety; patient outcomes in end-stage cancer - the MetAction study.

Background: In precision cancer medicine, the challenge is to prioritize DNA driver events, account for resistance markers, and procure sufficient information for treatment that maintains patient safety. The MetAction project, exploring how tumor molecular vulnerabilities predict therapy response, first established the required workflow for DNA sequencing and data interpretation (2014-2015). Here, we employed it to identify molecularly matched therapy and recorded outcome in end-stage cancer (2016-2019).Material and methods: Metastatic tissue from 26 patients (16 colorectal cancer cases) was sequenced by the Oncomine assay. The study tumor boards interpreted called variants with respect to sensitivity or resistance to matched therapy and recommended single-agent or combination treatment if considered tolerable. The primary endpoint was the rate of progression-free survival 1.3-fold longer than for the most recent systemic therapy. The objective response rate and overall survival were secondary endpoints.Results: Both common and rare actionable alterations were identified. Thirteen patients were found eligible for therapy following review of tumor sensitivity and resistance variants and patient tolerability. The interventions were inhibitors of ALK/ROS1-, BRAF-, EGFR-, FGFR-, mTOR-, PARP-, or PD-1-mediated signaling for 2-3 cases each. Among 10 patients who received treatment until radiologic evaluation, 6 (46% of the eligible cases) met the primary endpoint. Four colorectal cancer patients (15% of the total study cohort) had objective response. The only serious adverse event was a transient colitis, which appeared in 1 of the 2 patients given PD-1 inhibitor with complete response. Apart from those two, overall survival was similar for patients who did and did not receive study treatment.Conclusions: The systematic MetAction approach may point forward to a refined framework for how to interpret the complexity of sensitivity versus resistance and patient safety that resides in tumor sequence data, for the possibly improved outcome of precision cancer medicine in future studies. ClinicalTrials.gov, identifier: NCT02142036.

IL-29 Exhibits Anti-Tumor Effect on Pan-48 Pancreatic Cancer Cells by Up-regulation of P21 and Bax.

BACKGROUND/AIM: Pancreatic cancer is the most lethal cancer of the digestive system. IL-29 is a new member of the IFNλ family and well-known for its strong antiviral activity. However, its direct effect on pancreatic cancer is still unclear. This study was performed to investigate if IL-29 has any direct effect on Pan-48 pancreatic cancer cells. MATERIALS AND METHODS: Clonogenic survival assay, cell proliferation, and caspase-3 activity kits were used to evaluate the effects of IL-29 on cell survival, proliferation, and apoptosis of Pan-48 pancreatic cancer cells. RT-PCR and IHC were subsequently performed to explore IL-29's potential molecular mechanisms. RESULTS: The percentage of colonies of Pan-48 cells was decreased following the addition of IL-29. This was consistent with a decreased optical density (OD) value of cancer cells. Furthermore, the relative caspase-3 activity in cancer cells was increased after the addition of IL-29, indicating increased apoptosis of cancer cells. The anti-proliferative effect of IL-29 on cancer cells correlated with increased expression of the anti-proliferative molecule p21. The pro-apoptotic effect of IL-29 on cancer cells correlated with an increased expression of the pro-apoptotic molecule Bax. CONCLUSION: IL-29 constrains Pan-48 pancreatic cell growth via up-regulation of p21 and Bax. Our study suggests a potential use of IL-29 in immunotherapy for pancreatic cancer treatment.

p38 MAPK activation through B7-H3-mediated DUSP10 repression promotes chemoresistance.

Immunoregulatory protein B7-H3 is involved in the oncogenic and metastatic potential of cancer cells, as well as in drug resistance. Resistance to conventional chemotherapy is an important aspect of melanoma treatment, and a better understanding of how B7-H3 enhances drug resistance may lead to the development of more effective therapies. We investigated the in vitro and in vivo sensitivity of chemotherapeutic agents dacarbazine (DTIC) and cisplatin in sensitive and drug resistant melanoma cells with knockdown expression of B7-H3. We found that knockdown of B7-H3 increased in vitro and in vivo sensitivity of melanoma cells to the chemotherapeutic agents dacarbazine (DTIC) and cisplatin, in parallel with a decrease in p38 MAPK phosphorylation. Importantly, in B7-H3 knockdown cells we observed an increase in the expression of dual-specific MAP kinase phosphatase (MKP) DUSP10, a MKP known to dephosphorylate and inactivate p38 MAPK. DUSP10 knockdown by siRNA resulted in a reversion of the increased DTIC-sensitivity seen in B7-H3 knockdown cells. Our findings highlight the potential therapeutic benefit of combining chemotherapy with B7-H3 inhibition, and indicate that B7-H3 mediated chemoresistance in melanoma cells is driven through a mechanism involving DUSP10-mediated inactivation of p38 MAPK.

Correction to: IL-39 acts as a friend to pancreatic cancer.

The original version of this article unfortunately contained a mistake in the text of the entire article. The word "IL-39" should read as "meteorin-like protein". This has been corrected with this correction.

IL-39 acts as a friend to pancreatic cancer.

Pancreatic cancer is the most lethal digestive cancer and the fourth leading cause of cancer death in the US. IL-39, a heterodimer of p19 and EBI3, is a newly found cytokine and its role in the pathogenesis of neoplasia has not been studied yet. This study was designed to investigate the direct role of IL-39 in the growth of pancreatic cancer. Clonogenic survival assay, cell proliferation, and caspase-3 activity kits were used to evaluate the direct effects of IL-39 on cell survival, proliferation and apoptosis of the widely studied pancreatic cancer cell line MiaPaCa-2. We further investigated the possible molecular mechanisms by using RT-PCR and IHC. The percentage of colonies of pancreatic cancer cells increased significantly in the presence of IL-39. This was paralleled with the increase in the OD value of cancer cells in the presence of IL-39. Interestingly, the relative caspase-3 activity in cancer cells decreased significantly in the presence of IL-39. Furthermore, the pro-tumor effect of IL-39 on pancreatic cancer cells correlated with decreased anti-proliferative molecule p21.The anti-apoptotic effect of IL-39 correlated with decreased pro-apoptotic molecule TRAILR1. These results suggest that IL-39 favors growth of pancreatic cancer by promoting growth and inhibiting apoptosis of cancer cells. This suggests that IL-39 acts as a friend to pancreatic cancer. Thus, inhibition of effect of IL-39 on cells might be a promising strategy to treat pancreatic cancer.

The novel role of IL-37 in prostate cancer: evidence as a promising radiosensitizer.

Prostate cancer (PCa) is the most common non-cutaneous cancer in men in the USA. Radiation therapy (RT) is widely considered the standard treatment for PCa. IL-37 is an IL-1 family member, and it has been extensively studied in immunity. However, no studies have been done regarding its potential as a radiosensitizer. This study is designed to investigate the direct effect of IL-37 on growth of DU145 and PC-3, two widely studied PCa cell lines, and to investigate whether IL-37 could be used as a radiosensitizer for PCa. Clonogenic survival and quick cell proliferation assays along with immunohistochemistry, TUNEL staining, and caspace-3 activity assay kits as well as RT-PCR were used in this study. Our results showed that IL-37 has little direct effect on growth of PCa. However, IL-37/RT enhanced RT-induced inhibition of cell proliferation and apoptosis in both cell lines. We further found that IL-37/RT upregulated the mRNA expression of p27, Fas, and Bax, while downregulating the mRNA expression of cdk2 in DU145 cells. These findings suggest that IL-37 has the potential to be used as a radiosensitizer for PCa and warrants further investigation.

Unveil the mysterious mask of cytokine-based immunotherapy for melanoma.

Melanoma is the leading cause of death among all skin cancers and its incidence continues to rise rapidly worldwide in the past decades. The available treatment options for melanoma remain limited despite extensive clinical research. Melanoma is an immunogenic tumor and great advances in immunology in recent decades allow for the development of immunotherapeutic agents against melanoma. In recent years, immunotherapy utilizing cytokines has been particularly successful in certain cancers and holds promise for patients with advanced melanoma. In this review, an overview of the current status and emerging perspectives on cytokine immunotherapy for melanoma are discussed in details. Such a study will be helpful to unveil the mysterious mask of cytokine-based immunotherapy for melanoma.

The role of IL-7 in Immunity and Cancer.

Interleukin-7 (IL-7) is a cytokine that has been known since long in immunology, mainly regarding its effects on T-cells and B-cells. IL-7 has been demonstrated to be necessary for both B-cell and T-cell proliferation and lack of IL-7 causes immature immune cell arrest. Interestingly, in recent years, certain studies have strongly suggested that the role of IL-7 is far beyond the field of immunology, it might have direct or indirect effect on cancer. This review aims to summarize the role of IL-7 in immunity and its role in the pathogenesis of neoplasia.

The Role of IL-37 in Non-Cancerous Diseases.

IL-37 is a newly discovered cytokine belonging to IL-1 family consisting of 11 members, which have similar ß-barrel structures and associate with Ig-like receptors. Extensive studies have been done with IL-37 since its discovery. These studies suggest that IL-37 does not only play a role in tumorigenesis, but also has anti-inflammatory properties in immune responses through the down regulation of pro-inflammatory molecules. We have previously reviewed the role of IL-37 in cancer. Here, we will focus on the role of IL-37 in non-cancerous Diseases. Such a study might be helpful to design new strategies to treat IL-37 associated diseases.

The role of IL-29 in immunity and cancer.

Interleukin-29 (IL-29) is a new member of the recently discovered interferon λ (IFNλ) family. It is produced predominantly by maturing dendritic cells and macrophages. It has been implicated in numerous immunological responses and has shown antiviral activity similar to the Type I interferons, although its target cell population is more limited than the Type I interferons. In recent years, the role of IL-29 in the pathogenesis of various cancers has also been extensively studied. In this review, we will discuss the recent advances of IL-29 in immunological processes and the pathogenesis of various cancer.

The role of IL-37 in cancer.

Interleukin 37 (IL-37) is a new member of the IL-1 family which all have a similar ß-barrel structure. Since its discovery, IL-37 has been studied extensively in immunological field. It has been established that IL-37 possesses anti-inflammatory characteristics both in innate immune response as well as in acquired immune responses by downregulating pro-inflammatory molecules. This review will discuss the role of IL-37 in immunological processes and neoplastic pathogenesis.

Proton nuclear magnetic resonance spectroscopy of urine samples in preterm asphyctic newborn: a metabolomic approach.

In order to highlight differences in the metabolic profile of healthy (control) compared with asphyxiated newborns, by using untargeted metabolomic approach coupled with (1)H NMR spectroscopy, we evaluated the effects of asphyxia on newborn urine metabolites. Our results showed that lactate, glucose and TMAO, together with threonine plus 3-hydroxyisovalerate are the metabolites more characterizing the asphyxiated group; lower contribute to discrimination is related to other metabolites such as dimethylglycine, dimethylamine, creatine, succinate, formate, urea and aconitate. After 24-48h from resuscitation preterm asphyctic neonates showed their recovery pattern that still can be differentiated by the controls.

Using metabolomics to monitor kidney transplantation patients by means of clustering to spot anomalous patient behavior.

BACKGROUND: NMR spectroscopy-based metabolomics is a system approach used to investigate the metabolic profile of biological fluids with multivariate data analysis tools. The aim of this study was to examine the kidney graft recovery process noninvasively through the examinations of urine samples using (1)H NMR spectroscopy combined with chemometric tools. METHODS: Urine samples were treated as the source of metabolites reflecting the pathological and clinical conditions of patients with transplanted kidneys. We observed 15 subjects (9 males and 6 females) during the graft recovery process and initial days thereafter. The patients provided at least 9 samples each, applying advanced statistical methods of analysis: Principal Component Analysis (PCA) and Partial Least Square Discriminant Analysis PLS-DA). RESULTS: The PCA model (for all subjects exp. var. PC1 13.96% and PC2 9.88%) allowed us to clearly designate 3 stages of recovery: initially the kidney is not working; in the second stage, it regains functions, and the third stage includes follow-up during hospitalization. PCA analysis of a single patient follows graft recovery based on biochemical (metabolites) information, assigning the appropriate recuperation stage. CONCLUSIONS: NMR spectroscopy together with chemometric analysis allow monitoring of kidney graft recovery to identify patients who are not progressing within the normal range.

AlkB demethylases flip out in different ways.

Aberrant methylations in DNA are repaired by base excision repair (BER) and direct repair by a methyltransferase or by an oxidative demethylase of the AlkB type. Yang et al. [Nature 452 (2008) 961-966] have now solved the crystal structure of AlkB and human AlkB homolog 2 (hABH2) in complex with DNA using an ingenious crosslinking strategy to stabilize the DNA-protein complex. AlkB proteins have similar catalytic domains, but different DNA recognition motifs. Whereas AlkB mainly makes contact with the damaged strand, hABH2 makes numerous contacts with both strands. hABH2 flips out the damaged base and fills the vacant space by a hydrophobic amino acid residue similar to DNA glycosylases, essentially without distorting the double helix structure. In contrast, AlkB squeezes together the bases flanking the flipped-out base to maintain the base stack. This unprecedented flipping mechanism and the differences between AlkB and hABH2 in contacting the DNA strands explain their preferences for single stranded- and double stranded DNA, respectively.

Ets-1 transcription factor is widely expressed in benign and malignant melanocytes and its expression has no significant association with prognosis.

Ets-1 transcription factor has been associated with tumor progression in various carcinomas, but its expression in malignant melanoma was only recently described. The study was conducted in two steps: exploratory and confirmatory. In the first step, we studied 69 primary melanomas, 28 metastatic melanomas, 10 usual intradermal nevi and 13 various melanocytic skin lesions. In the second step, an additional group of 98 patients with follow-up of up to 200 months was also evaluated. Immunohistochemical analysis of formalin-fixed/paraffin-embedded tissues was performed using 1G11 antibody and polymer conjugate for visualization. While Ets-1 was variably expressed in 83% primary melanomas in exploratory and 69% in the confirmatory group, the expression of Ets-1 was also found in normal benign melanocytes and all nevi. Analysis of the exploratory group revealed lower expression of Ets-1 in primary melanomas than in common nevi (P=0.048, Mann-Whitney U-test) and metastatic melanomas expressed significantly less Ets-1 than primary melanomas (P=0.015, Mann-Whitney U-test). There was a negative correlation between Ets-1 expression and the largest dimension of the primary tumors (r=0.23, P=0.034, Spearman's correlation rank test), but no correlation with the depth of tumor invasion (Breslow thickness) or the presence of ulceration was found. Analyses of the confirmatory group revealed no association between Ets-1 expression with disease-specific survival or time to treatment failure. However, a statistical trend was found for worse outcome for those primary melanomas that had strong expression (H-score >100) of Ets-1 (P=0.054). Ets-1 is expressed in benign melanocytes probably due to their neural crest origin. We conclude that Ets-1 expression cannot be used to differentiate between benign and malignant melanocytic lesions and it has no definite association with clinical outcome. At the same time, its role in tumor progression in some cases of malignant melanoma cannot be entirely excluded.

Expression of S100A4 combined with reduced E-cadherin expression predicts patient outcome in malignant melanoma.

The aim of the present study was to analyze the expression of S100A4 and E-cadherin in a panel of primary and metastatic malignant melanoma, and to correlate the expression level to clinicopathological parameters. The expression of S100A4 was examined by immunohistochemistry in 99 superficial spreading and 60 nodular primary melanomas, while the expression of E-cadherin was analyzed in 92 superficial spreading and 52 nodular lesions from the same panel. The expression levels of S100A4 and E-cadherin in the biopsies were inversely correlated, with S100A4 being expressed at the highest frequency in the nodular and E-cadherin in the superficial spreading lesions, respectively. When analyzing the melanoma subgroups separately, it was revealed that expression of S100A4 had a more significant impact on patient outcome in early superficial spreading melanomas than in the nodular subtype, while E-cadherin expression did not predict patient outcome in any of the subgroups. When examining all the patients, both markers give clinical information as predictors for disease-free survival, but when combining the expression of the two markers, a stronger significant correlation between high E-cadherin expressing/S100A4 negative biopsies and increased disease-free survival (P=0.002) was revealed, demonstrating the importance of examining the expression of more than one factor involved in the metastatic cascade when predicting patient outcome. We have also evaluated the relationship between the expression of these two antigens and cell cycle and signal transduction factors.

Clinical and cell line specific expression profiles of a human gene identified in experimental central nervous system metastases.

Some cancers, particularly malignant melanomas and carcinomas of the breast and lung, metastasize to the central nervous system (CNS) in advanced stages. In order to develop into clinically manifest metastases, hematogenously disseminated tumor cells must respond to trophic factors within the CNS microenvironment. We have previously identified a nuclearfactor, com1, expressed in human breast carcinoma cells upon formation of experimental metastatic tumors in the CNS. In the present study distinct com1 mRNA expression was detected in cerebral metastases from patients with lung carcinomas, whereas the expression level was generally much lower in glioblastomas (primary brain tumors). In tissue specimens from normal brain and lung, as well as in glioma and lung carcinoma cell lines, com1 expression was barely detectable. One potential mechanism involved in the induction of com1 expression was indicated in the metastatic MCF7/LCC2 breast carcinoma cells. Significant increases in the level of com1 mRNA were observed upon activation of receptor tyrosine kinase signaling, which is known to operate during metastatic tumor cell proliferation within the CNS. The observations in this study strengthen the assumption that com1 may be involved in the tumor cell response to regulatory signals upon metastasis formation.

NMR spectroscopic characterization of sarcolemmal permeability during myocardial ischemia and reperfusion.

This study aims to characterize the pattern of membrane disintegration during myocardial ischemia and reperfusion. Intracellular volumes were measured by 1H and 59Co NMR in isolated rat hearts during 10, 30 and 60 min of total ischemia and 30 min of reperfusion at normothermia. Perfusion with hypo-osmotic medium (210 mosm/l) increased intracellular water from 2.50+/-0.06 to 3.07+/-0.07 ml/g dry weight (P<0.001) during pre-ischemia. Hypo-osmotic swelling decreased by 16+/-3, 32+/-6 and 44+/-11% of the pre-ischemic value after 10, 30 and 60 min of ischemia (n.s., P<0.005, P<0.001) respectively, indicating that membrane permeabilization facilitated efflux of osmolytes and counterbalanced the osmotic driving force for water influx. Hypo-osmotic swelling decreased during 30 min of reperfusion by 18+/-5% in all groups (P<0.0.005 v post-ischemia). The volume of distribution of the extracellular marker cobalticyanide increased by more than 3.2+/-0.4 and 5.8+/-0.5% of the intracellular space after 30 and 60 min of ischemia respectively (P<0.001), and by an additional 2% after reperfusion. During 30 min of reperfusion, hearts released 1.6+/-0.2 and 3.2+/-0.4% of the intracellular creatine kinase contents after 30 and 60 min of ischemia, respectively (P<0.001). In addition to the correlation between ischemia duration and membrane permeability, evident from the analysis of each probe, the data showed a progressive increase in severity of membrane injury over time and permeabilization to larger molecules. 23Na NMR spectroscopy in conjunction with an extracellular shift reagent (SR) showed formation of a resonance at an intermediate chemical shift in between the intra and extracellular Na+ peaks, suggesting penetration of SR into cells with disrupted membranes. The constant chemical shift and narrow line shape of this resonance, characteristic of a homogeneous chemical environment, suggested that the distribution of SR was contained within the cytosol of cardiomyocytes. We propose that sarcolemmal membranes are gradually permeabilized to larger molecules by ischemia, and the evolving chemical instability is spatially contained within the myocyte.