Dead Mice Research Articles

The Effect of Xylazine/Zoletil Anesthesia on the Radiosensitivity of Mice under Total Irradiation with X-Rays, Protons, and Carbon Ions.

The effect of xylazine-zoletyl anesthesia on the radiosensitivity of mice irradiated with protons, carbon ions in two regions of the Bragg curve, and X-ray radiation was studied according to the criteria of 30-day survival, dynamics of death, and the average life span of dead mice. The maximum effect of anesthesia by 3.3 times was observed by a decrease in the death of animals during irradiation with carbon ions at the Bragg peak; in the case of irradiation before the peak, the effect was 1.2 times. In the case of proton irradiation at the Bragg peak, the protective effect of anesthesia by a factor of 1.7 was observed only at a dose of 8.5 Gy. When mice were irradiated with X-rays in the dose range of 6.0-8.5 Gy, the anesthesia effect coefficient was 1.7-2. According to the 30-day survival method, it was shown that the use of a xylazine-zoletil mixture significantly changes the radiosensitivity of mice depending on the radiation dose and the radiation source quality.

COMPLETE UNILATERAL MAXILLECTOMY IN A COHORT OF FIVE COLOMBIAN BOAS (BOA IMPERATOR).

Five unrelated adult Colombian boas (Boa imperator) presented with a 1- to 3-mon history of unilateral rostral swelling of the maxilla associated with a chronic rubbing against the enclosure's walls. Moderate to severe gingival inflammation and ulceration of the labial mucosa were present at the level of the swelling with tenderness to the touch. Radiography revealed osteolytic or proliferative lesions of the maxillary bone. Chronic maxillary osteomyelitis was diagnosed. Unilateral maxillectomy was performed on each animal under general anesthesia. Local anesthesia was also achieved by infiltrating lidocaine along the medial and lateral aspect of the maxillary gingiva and at the level of the maxillo-ectopterygoid joint. Using a lateral gingival approach, the maxillo-prefrontal, maxillary-palatine, and maxillo-ectopterygoid attachments were transected, and the maxillary bone removed. Histologic examination revealed pyogranulomatous stomatitis and osteomyelitis in all snakes, and presence of intralesional bacteria (n = 3 snakes). Gram-negative bacteria (Chryseobacterium indologenes and Proteus mirabilis) were cultured from the resected tissue of two snakes. One snake suffered from wound dehiscence 5 d postoperatively. All snakes were fed 15 d postoperatively and ingested dead mice without apparent difficulties. One snake was examined 2 mon and 1 yr after surgery, with no evidence of soft tissue or osseous infection and only minor facial scaring; all other snakes were lost to follow-up 15 d after surgery. Unilateral maxillectomy was performed in a cohort of five Colombian boas suffering from maxillary osteomyelitis. This surgical technique should be considered as an alternative to medical treatment in boid snakes.

Transmembrane TNF-α as a Novel Biomarker for the Diagnosis of Cytokine Storms in a Mouse Model of Multiple Organ Failure

A cytokine storm (CS) is an out-of-control inflammatory response closely associated with the progression of diseases, such as multiple organ failure (MOF), severe sepsis, and severe or critical COVID-19. However, there is currently a lack of reliable diagnostic markers to distinguish CS from normal inflammatory responses. Tumor necrosis factor-α (TNF-α) includes transmembrane TNF-α (tmTNF-α) and secreted TNF-α (sTNF-α). The MOF mouse model in this study showed that the tmTNF-α expression changes in the neutrophils differed from the serum TNF-α and serum IL-18, INF-γ, IL-4, and IL-6. Furthermore, tmTNF-α, instead of serum TNF-α, IL-18, INF-γ, IL-4, and IL-6, reflected liver and kidney tissue damage and increased with the aggravation of these injuries. Analysis of the ROC results showed that tmTNF-α effectively distinguished between inflammatory responses and CS and efficiently differentiated between surviving and dead mice. It also significantly improved the diagnostic value of the traditional CRP marker for CS. These results indicated that the tmTNF-α expressed in the neutrophils could be used to diagnose CS in MOF mice, providing an experimental basis to further develop tmTNF-α for diagnosing CS patients.

On Felt, Mice and Moths Change and Permanence in a Work by Joseph Beuys


 
 
 
 
 In November 1970, Joseph Beuys and Terry Fox performed Isolation Unit in Düsseldorf in the cellar of the Kunstakademie, an hour-long performance in front of an audience of about thirty people. As in many of Beuys’s performances, the relation with (live or dead) animals and the use of natural materials played a crucial role. This is evident by the title he later used for the work: Action the dead mouse. During this performance, Beuys wore, for the first time, the felt suit that would later become one of his most iconic art-objects. Felt, like fat, is one of those organic matters recurring in his actions and refers to the warmth of the body, the energy of life. Shortly after the performance, the Felt Suit was (re)produced as a multiple in an edition of 100 copies by the Galerie René Block in Berlin. This essay starts with Isolation Unit and analyzes strategies for dealing with the organic and inorganic transformations of the multiple that originated from it. In doing so, it investigates contemporary approaches to the preservation of ‘ephemeral’ works and performance
 
 
 
 
 
 
 remains and discusses their consequences for the epistemology and ontology of the arts.
 
 
 
 

Effect of Interleukin-6 Gene Deletion on Radiation-Induced Mouse Hematopoietic Injury and Relative Mechanism

To study the effect of interleukin-6 (IL-6) gene deletion on radiation-induced hematopoietic injury in mice and relative mechanism. Before and after whole body 60Co γ-ray irradiation, it was analyzed and compared that the difference of peripheral hemogram, bone marrow hematopoietic stem and progenitor cells conts in IL-6 gene knockout (IL-6-/-) and wild-type (IL-6+/+) mice and serum IL-6 and G-CSF expression levels in above- mentioned mouse were detected. Moreover, 30 days survival rate of IL-6-/- and IL-6+/+ mice after 8.0 Gy γ-ray irradiation were analyzed. IL-6 levels in serum of IL-6+/+ and IL-6-/- mice were respectively (98.95±3.85) pg/ml and (18.36±5.61) pg/ml, which showed a significant statistical differences (P<0.001). There were no significant differences of peripheral blood cell counts and G-CSF level in serum between IL-6+/+ and IL-6-/- mice before irradiation (P>0.05). However, the number of leukocytes, neutrophils, lymphocytes, monocytes, platelets in peripheral blood and G-CSF level in serum of IL-6-/- mice were significantly decreased at 6 h after 8.0 Gy γ-ray irradiation compared with that of IL-6+/+ mice. On days 30 after 8.0 Gy γ-ray irradiation, the survival rate of IL-6+/+ and IL-6-/- mice was 62.5% and 12.5%, and the mean survival time of dead mice was 16.0±1.0 and 10.6±5.3 days, respectively. On days 14 after 6.5 Gy γ-ray irradiation, bone marrow nucleated cells in IL-6+/+ and IL-6-/- mice were respectively (10.0±1.2)×106 and (8.3±2.2)×106 per femur. Compared with IL-6+/+ mice, the proportion of Lin-Sca-1-c-kit+ (LK) in bone marrow of IL-6-/- mice had no significant change (P>0.05), but the proportion of Lin-Sca-1+c-kit+ (LSK) was significantly decreased (P<0.05). IL-6 plays an obvious role in regulating hematopoietic radiation injury, and IL-6 deficiency can inhibit the radiation-induced increase of endogenous G-CSF level in serum, aggravates the damage of mouse hematopoietic stem cells（HSC） and the reduction of mature blood cells in peripheral blood caused by ionizing irradiation, resulting in the shortening of the survival time and significant decrease of the survival rate of mice exposed to lethal dose radiation.

Study on the hypoglycemic effect of mango leaf extract grown in Son La

Mango leaves were extracted by Soxhlet with 60% ethanol, the obtained solution was distilled to remove the solvent under reduced pressure. The results showed that when using the maximum allowable dose, there were no dead mice. Experimenting on blood glucose-lowering effects on diabetic white mice obtained the results: with a dose of 500 mg/kg giving the best therapeutic effect, even slightly better than the positive control drug gliclazide, the blood glucose level decreased immediately after three days. After 21 days reduced to 52.5% and brought the blood glucose index to normal. This is a new direction in the use of drugs of plant origin in general and Mango leaves in particular.

The Probiotic Attributes and Anti-pseudorabies Virus Evaluation of Lactobacillus Isolates.

The emergence of pseudorabies virus (PRV) variants brings serious harm to the swine industry, and its effective treatments are limited at present. As one of the probiotics, the Lactobacillus species have beneficial characteristics of regulating the balance of intestinal flora, inhibiting the growth of pathogenic bacteria and viruses' proliferation, and improving self-immunity. In this study, Lactobacillus plantarum HN-11 and Lactobacillus casei HN-12 were selected and identified through morphology observation, Gram stain microscopy, 16S rRNA sequencing analysis, and specific amplification of the recA gene and pheS gene. All tested isolates exhibited rapid adaptation to the different conditions, excellent acid, and bile tolerance, and sensitivity to Salmonella, Staphylococcus aureus, and Escherichia coli. The antibiotic susceptibility assay displayed the isolates sensitive to most antibiotics and resistant to Lincomycin and Norfloxacin. Moreover, the supernatants of HN-11 and HN-12 inhibited PRV proliferation in ST cells. The results of animal experiments showed that supplementing the challenged mice with the supernatants of Lactobacillus isolates in advance delayed the course of the disease. PRV was detected in the heart, liver, spleen, lung, kidney, and brain tissues of dead mice in the test groups, and its copies in the lungs were significantly decreased compared with the control mice (P < 0.05). These findings proved the advantages of L. plantarum and L. casei as potential probiotic cultures, which could provide a basis for its application in microecological preparations and functional formulations.

Abstract 2122: CRD1600 is a potent HPK-I inhibitor with robust immune modulatory properties

Abstract Background: Haematopoietic Progenitor Kinase (HPK-1, MAP4K1) is a member of the STE20 family of serine/threonine kinases, expressed predominantly in hematopoietic cells. The kinase activity of HPK-1 provides an inhibitory signal that dampens immune responses1. Upon activation of T-cell and/or B-cell receptors, activated HPK-1 phosphorylates the adaptor protein SLP76 at Ser376 in T cells or BLNK at Thr152 in B-cells to trigger proteasomal degradation of the adaptors and disrupt downstream T-cell or B-cell activating signalosome complexes. Blocking HPK-1 extends immune action and since HPK-1 expression is primarily restricted to immune cells, it is a compelling drug target for immune modulation. Anti-tumor efficacy has been demonstrated in HPK-1 kinase dead knock-in mice, HPK-1 knockout mice or using small molecule pharmacological inhibitors of HPK-1 either as monotherapy or in combination regimens2. Methods: Curadev used a pharmacophore based approach to discover novel small molecule HPK-1 inhibitors. SAR studies were carried out using a screening funnel that included efficacy assessment by monitoring in vitro enzymatic and cellular activity, anti-target cross-kinase specificity by assessing cytotoxicity in fresh murine splenocytes and effector mobilization by monitoring IL2 production and human T-cell proliferation under anti-CD3/CD28 stimulation. Results: Curadev’s medicinal chemistry campaign yielded several highly potent HPK-1 inhibitors with sub-nanomolar to nanomolar potency in in vitro human HPK-1 enzyme assays which were active in the cellular SLP76 assay. A single dose of the lead compound CRD1600 to mice treated with an anti-CD3 antibody caused systemic enhancement of pro-inflammatory cytokines such as IL2 and reduced pSLP76 levels in the spleen. These pharmacodynamic effects correlated with plasma levels of CRD1600. Conclusions: Inhibition of HPK-1 is a promising therapeutic modality that could augment the effects of existing anti-cancer treatments including immunotherapy. CRD1600 is a potent, selective HPK-1 inhibitor that has been selected from a rich portfolio of active compounds and shows in vivo activity against the target. Further development of the compound is ongoing.

Acute toxicity of nano Berberine gel in white mice.

Berberine is isolated from Coscinium fenestratum and some plants in the family Ranunculaceae which has the effect of treating burn wounds. The study determined acute oral toxicity administration in white mice of nano Berberine gel (NBBR) produced by the Le Huu Trac National Burns Hospital.Research methods: The research method was accorded to the guidelines of the Ministry of Health, World Health Organization. Evaluation of 120 mice, divided into 10 groups, NBBR dosages were gradually increased from 30.9g/kg to 123.96g/kg by oral administration. Results: No dead mice, no abnormal disorders in movement and digestion; no abnormal histopathological found in livers, spleens and kidneys.Conclusion: LD50 (Median Lethal Dosage) of NBBR gel had not been determined in white mice by oral administration with the highest dose that could be given 123.96g/kg body weight/day.

PEG-grafted liposomes for enhanced antibacterial and antibiotic activities: An in vivo study

Staphylococcus aureus (S. aureus) biofilm-associated infections are a primary concern for public health worldwide. Current therapeutics cannot penetrate the biofilms efficiently, resulting in low drug concentrations at the infected sites and increasing the frequency of drug usage. To solve this issue, nanotechnology platforms seem to be a promising approach. In this study, the potential therapeutic effects of (PEG)ylated liposome (PEG-Lip) for the delivery of nafcillin (NF) antibiotic were assessed. The results demonstrated that NF-loaded liposome (Lip-NF) and NF-loaded PEG-Lip (PEG-Lip-NF) released 76.4 and 62% of the loaded NF, respectively, in a controlled manner after 50 h. Also, it was found that PEG-Lip-NF, compared to Lip-NF and NF, was more effective against a methicillin-susceptible S. aureus (MSSA; minimum inhibitory concentration (MIC): 1.0 ± 0.03, 0.5 ± 0.02, and 0.25 ± 0.01 μg/mL; and minimum biofilm inhibitory concentration (MBIC50): 4.0 ± 0.18, 1.0 ± 0.04, and 0.5 ± 0.02 μg/mL for NF, Lip-NF, and PEG-Lip-NF, respectively). PEG-Lip-NF, compared to NF and Lip-NF, could also more efficiently decrease the side effects of NF through improving human MG-63 osteoblast cell viability (cell viability at 100 μM of NF: 76, 68, and 38% for PEG-Lip-NF, Lip-NF, and NF, respectively). PEG-Lip-NF, compared to control, NF, and Lip-NF groups, was more efficacious by 45, 25, and 10%, respectively, to decrease the virulence of MSSA bacteremia through inhibiting the weight loss of the infected mice. Also, PEG-Lip-NF and Lip-NF, compared to control and NF groups, caused a considerable decrease in the mortality rate in a murine model of bacteremia (number of dead mice: 0, 0, 2, and 8 out of 15 for PEG-Lip-NF, Lip-NF, NF, and control groups, respectively). Overall, the results of this study demonstrated that the loading of NF into PEG-Lip is a promising strategy to decrease the side effects of NF with improved antibacterial effects for the treatment of MSSA biofilm-associated infections.

MLN0128, a Potential Target Therapy for Myeloid Proliferation Neoplasia (MPN) with a Benefit of Prolonging Survival

MLN0128 (TAK-228, INK128) is an investigational new drug (IND) that is currently being evaluated in clinical trials and has showed promising efficacies in patients with solid cancer. MLN0128 is one of the second generation of inhibitors for mammalian target of rapamycin (mTORi), which suppresses the dysregulated cell survival signaling activities in cancer cells by specifically inhibiting both mTORC1 and mTORC2 in PI3K/mTOR pathway. MLN0128 was reported inhibiting acute myeloid leukemia (AML) stem cells and progenitors [Zeng et al, Oncotarget, 2016]. However, it is yet to be clarified whether MLN0128 can improve survival time in leukemia patients. Here, we report data from the investigation of the impact of MLN0128 on leukemia survival in mice. We tested MLN0128 in a published mouse model which simultaneously produces experimental mice with genotypes that are associated with pediatric leukemia, including myeloproliferative neoplasm (MPN) and juvenile myelomonocytic leukemia (JMML), and adult chronic myelomonocytic leukemia (CMML) [Vara et al, Blood Adv.2020]. Specifically, mice with deletion of Phosphatase and tensin homolog gene (Pten) and loss-of-heterozygosity of Neurofibromin 1 (Nf1) (Pten Δ/ΔNf1 LOH) develop JMML, mice with Pten deletion and Nf1 heterozygosity (Pten Δ/ΔNf1 Δ/wt) develop pediatric MPN, both of which cause juvenile death; mice with a single copy of Pten deletion and Nf1 LOH (Pten Δ/wtNf1 LOH) develop CMML in adulthood. We previously reported that all of those diseased mice shared a feature with elevated activities in PI3K/MAPK/mTOR pathways. We first collected nucleated cells from bone marrow (BM) of the diseased mice and tested the inhibition of MLN0128 on colony formation of hematopoietic progenitors (CFU-GM) at a concentration range of 1-1000nM. Our data demonstrated that MLN028 significantly inhibited the CFU-GM colony formation of BM cells from mice with all three diseased genotypes, in a dose-dependent fashion. Importantly, the IC50 for CFU-GM was less than 100nM, which is in the concentration range of MLN0128 in the blood from patients who orally received MLN0128 at dosage range of 4-12mg/day [Ghobrial et al AJH 2016]. We then investigated the impact of MLN0128 on life span in diseased mice. We administrated MLN0128 in gavage at dosages of 0.5-1μg/g body weight, starting at an age of postnatal day 17-18 (PND17-18), 3-5 times a week, repeated for 5 weeks followed by a pause in 6 th week every course. Our data showed that MLN0128 significantly reduced the spleen sizes in treated mice with all diseased genotypes comparing to those treated with vehicle. The survival time was significantly prolonged in MLN0128-treated mice with genotypes of MNP (n=7 vs 3, median 150 vs 102 days) and CMML (n=7 vs 6, median 100 vs 56 days), comparing with the mice treated by vehicle. Unfortunately, we did not observe a survival improvement in treated JMML mice, but substantially swollen kidneys were found in the dead JMML mice, which was likely due to tumor-lysis syndrome because JMML mice carried a significantly worse leukemic burden. In conclusion, our data demonstrate that MLN0128 can significantly inhibit colony formation of BM cells from mice with genotypes of JMML, MPN, and CMML; and MLN0128 can significantly expand survival time of mice with genotypes of MPN and CMML, which has never been reported in previous studies that were related to mTORi in pediatric leukemia. Our data suggest that MLN0128 may be a potential target therapy for pediatric MPN and CMML with a benefit of prolonging survival. Further investigation is ongoing to evaluate the potential of MLN0128 in improving survival of JMML mice after minimizing the tumor-lysis syndrome by tipping down MLN0128 dosage. DisclosuresBertaina: Cellevolve Bio: Membership on an entity's Board of Directors or advisory committees; Neovii: Membership on an entity's Board of Directors or advisory committees; AdicetBio: Membership on an entity's Board of Directors or advisory committees.

Deterring non-target birds from toxic bait sites for wild pigs

Toxic baiting of wild pigs (Sus scrofa) is a potential new tool for population control and damage reduction in the US. Field trials testing a prototype toxic bait (HOGGONE 2 containing 5% sodium nitrite [SN]), though, revealed that wild pigs spilled small particles of toxic bait outside of bait stations which subsequently created hazards for non-target species that consumed those particles, primarily passerine birds. To deter non-target birds from consuming particles of spilled bait, we tested four deterrents at mock bait sites (i.e., baited with bird seed) in north-central Colorado, USA during April–May 2020. We found a programable, inflatable deterrent device (scare dancer) reduced bird visitation by an average of 96%. Then, we evaluated the deterrent devices at SN-toxic bait sites in north-central Texas, USA during July 2020, where the devices were activated the morning following deployment of SN-toxic bait. Overall, we found 139 dead wild pigs at 10 bait sites following one night of toxic baiting, which represented an average of 91% reduction in wild pigs visiting bait sites. We found that deterrent devices were 100% effective at deterring birds from toxic bait sites. We found two dead non-target mice at bait sites without deterrent devices. We noted that deploying toxic bait in mid-summer rather than late-winter/early-spring reduced hazards to migrating birds because they were not present in our study area during July. We recommend using deterrent devices (i.e., novel, programmable, battery operated, continuous and erratic movement, and snapping sounds) to reduce hazards to non-target birds at SN-toxic bait sites. We further recommend deploying SN-toxic bait during seasons when migrating birds are not as abundant until further research demonstrates minimal risks to migrating birds.

HANTAVIRUS PULMONARY SYNDROME: A RARE CASE REPORT

TOPIC: Chest Infections TYPE: Fellow Case Reports INTRODUCTION: Hantavirus pulmonary syndrome (HPS) is a rare but potentially life-threatening viral illness transmitted to humans from the infected urine, droppings or saliva of certain species of mice and rats. 728 cases have been reported so far since surveillance in the United States began in 1993 [1]. Most cases (93%) have occurred in states west of the Mississippi River. We present a case of a young male who presented with Hantavirus pulmonary syndrome. CASE PRESENTATION: 28 year old male from rural Texas presented with 1 week of intractable nausea and vomiting. He initially presented to an outside ER where he was found to be hypoxic requiring supplemental oxygen and worsening respiratory status and was transferred to our hospital for higher level of care. Initial workup with CT angiogram of chest revealed bilateral infiltrates and bilateral pleural effusions. Patient was initially started on empiric antibiotics and placed on non-invasive ventilation. Influenza and Repeated COVID-19 tests were negative. Patient reported that he lives in rural area and recently had exposure to a dead mouse in one of his house traps. Hantavirus IgG and IgM antibodies were sent which came back positive after patient had been discharged. Patient did well with supportive therapy of gentle diuresis and non-invasive ventilation and was discharged home after 5 days. DISCUSSION: The clinical syndrome of Hantavirus pulmonary syndrome (HPS) was first recognized in 1993. Although rare, it is frequently fatal, with a case fatality rate of 36%. HPS is an acute febrile illness with a prodrome consisting of fever, chills, myalgia, headache, and gastrointestinal symptoms. One or more of the following clinical features must be present:• Bilateral diffuse interstitial edema• Clinical diagnosis of acute respiratory distress syndrome (ARDS)• Radiographic evidence of noncardiogenic pulmonary edemaLaboratory criteria for diagnosis include detection of hantavirus-specific immunoglobulin M or rising titers of hantavirus-specific immunoglobulin G. CONCLUSIONS: Patients infected with Hantavirus pulmonary syndrome do better if they are recognized early and treated in Intensive care unit. There is no specific treatment or vaccine available at this point. Supplemental oxygen and intubation and mechanical ventilation for severe respiratory failure remains the standard of care. REFERENCE #1: https://www.cdc.gov/hantavirus/hps/index.html DISCLOSURES: No relevant relationships by Anam Hassan, source=Web Response No relevant relationships by Muhammad Hasham Sarwar, source=Web Response No relevant relationships by Brian Williams, source=Web Response

Differential pathogenesis of intracerebral and intramuscular inoculation of street rabies virus and CVS-11 strains in a mouse model.

Objective(s):The mechanisms of rabies evasion and immunological interactions with the host defense have not been completely elucidated. Here, we evaluated the dynamic changes in the number of astrocytes, microglial and neuronal cells in the brain following intramuscular (IM) and intracerebral (IC) inoculations of street rabies virus (SRV).Materials and Methods:The SRV isolated from a jackal and CVS-11 were used to establish infection in NMRI-female mice. The number of astrocytes (by expression of GFAP), microglial (by Iba1), and neuronal cells (by MAP-2) in the brain following IM and IC inoculations of SRV were evaluated by immunohistochemistry and H & E staining 7 to 30 days post-infection.Results:Increased numbers of astrocytes and microglial cells in dead mice infected by SRV via both IC and IM routes were recorded. The number of neuronal cells in surviving mice was decreased only in IC-infected mice, while in the dead group, this number was decreased by both routes.The risk of death in SRV-infected mice was approximately 3 times higher than in the CVS-11 group. In IC-inoculated mice, viral dilution was the only influential factor in mortality, while the type of strain demonstrated a significant impact on the mortality rate in IM inoculations.Conclusion:Our results suggested that microglial cells and their inflammatory cytokines may not contribute to the neuroprotection and recovery in surviving mice following intracerebral inoculation of SRV. An unexpected decrease in MAP2 expression via intramuscular inoculation indicates the imbalance in the integrity and stability of neuronal cytoskeleton which aggravates rabies infection.

Combinatorial Inhibition of Focal Adhesion Kinase and BCL-2 in AML

Focal adhesion kinase (FAK), activated by integrins and cytokines, promotes cell growth, migration and metastasis. We previously reported that a selective FAK inhibitor, VS-4718, inhibited cell growth and induced cell death in leukemia cell lines in vitro, effectively reduced leukemia burden in peripheral blood and tissues including bone marrow, spleen, liver and lung, and significantly prolonged survival time in a human xenograft murine model (Carter BZ et al., MCT, 2017), 16(6):1133).Although BCL-2 inhibition by the BCL-2 inhibitor ABT-199 (Venetoclax) has demonstrated antileukemia activities, an acquired drug resistance frequently ensues. When combined with ABT-199, VS-4718 significantly potentiated ABT-199-induced apoptosis in AML cell lines and primary AML cells. VS-4718 treatment of Molm14 AML cells decreased levels of p-STAT5, MCL-1, and BCL-XL (Wang et al., ASH 2016) that play important roles in the acquired resistance to ABT-199.To further examine the effect of VS-4718 as a single agent and in combination with ABT-199 on primary AML cells in vivo, we generated patient-derived xenografts (PDX) from AML patient bone marrow cells expanded in NSGS (NOD-SCID IL2Rgnull-3/GM/SF, NSG-SGM3) mice. The xenograft used AML cells from a patient who had failed multiple chemotherapies and had NPM1, FLT3-ITD, TET2, DNMT3A, and WT1 gene mutations and a complex karyotype. The combination of VS-4718 and ABT-199 synergistically killed the PDX cells in vitro even under MSC co-culture conditions. Once PDX cells were engrafted into NSGS mice, animals were randomized into 4 groups (N=10/group) and treated with vehicle, VS-4718 (75 mg/kg, twice daily), ABT-199 (100 mg/kg, once per day), or the combination via oral gavage for 29 days. We observed that ABT-199 had a stronger anti-leukemic effect on peripheral blood than on tissue-resident cells, while VS-4718 was more effective in tissues. At the end of the treatments, VS-4718 significantly reduced circulating human CD45+ cells (P=0.0003), and ABT-199 alone (P=5.90E-07) and the ABT-199/VS-4718 combination (P=5.38E-07) were ever more effective. VS-4718 alone or in combination with ABT-199 reduced leukemia burden in spleen and liver and effectively reduced splenomegaly, while ABT-199 alone did not. VS-4718-treated mice survived significantly longer than the untreated controls (median survival 72 vs. 36 days, P=0.0002), while ABT-199 did not prolong survival (median survival 35 days). Also, the combination of VS-4718 with ABT-199 significantly prolonged survival (median survival 65.5 days, P=0.011 vs. control and P=0.0072 vs. ABT-199). No statistical difference in survival was found between VS-4718 and the combination treatment groups (P=0.33) (Fig. 1). All the mice had large spleen at death except the combination group: the 1st dead mouse had small and the 2nd to 4th ones had slightly enlarged spleen and only the last two had large spleen as others suggesting that additional reasons such as toxicity at the dose used may have contributed to the death of the 1st 4 mice in the combination group, hence shortened the median survival in this group.We determined expression of FAK and BCL-2 family proteins in cells from mouse spleens at the end of treatment and found that the combination of VS-4718 and ABT-199 decreased protein levels of FAK. Consistent with our in vitro results, the increased level of MCL-1 by ABT-199 was antagonized by the combination with VS-4718 in vivo suggesting the combination potentially be more effective.Conclusions: These results suggest a potential clinical benefit of combining a FAK inhibitor with Venetoclax for patients with AML. [Display omitted] DisclosuresPachter:Verastem, Inc.: Employment. Weaver:Verastem, Inc.: Employment. Carter:Daiichi Sankyo: Research Funding; novartis: Research Funding.

ОСТРАЯ И ХРОНИЧЕСКАЯ ТОКСИЧНОСТЬ ЯНТАРНОЙ КИСЛОТЫ И ПРОБИОТИКА КПМ-2 ПРИ СОВМЕСТНОМ ИХ ПРИМЕНЕНИИ

The article discusses issues related to the determination of acute and chronic toxicity of succinic acid and probioic KPM-2 when used together in an experiment on white mice. A study of the indicators of acute toxicity showed that the experimental mice were marked by strong excitement, convulsions, death occurred from respiratory arrest during the period of convulsions. Autopsy of the dead mice observed hyperemia of the lungs, plethora of the liver, spleen, kidneys, and asphyxiation of the heart. Yak and probiotic KPM-2, when used together, do not possess cumulative properties according to the classification of Hodge and Sterner and belong to the 4th hazard class.

Adaptation of an artificial bait to an automated aerial delivery system for landscape-scale brown treesnake suppression

Dead neonatal mice are currently used as bait for delivery of toxin to invasive brown treesnakes (Boiga irregularis) on Guam; once deployed in the field the mice are highly attractive to the snakes but only for about four days. An artificial bait containing a mixture of fats mimicking those in skin of the mice is also highly attractive to the snakes and remains attractive 2–3 times longer. The artificial bait, however, costs more than the mice, and is more difficult to attach to the capsules of a novel aerial bait delivery system. This paper describes a reformulation of the bait which reduced the ingredient cost to 11% of its former value, without compromising its attractiveness to free-ranging snakes. Three lipid formulations using different source materials to produce the same fatty acid profile target were tested along with two levels of pork fat to yield six test formulations. No differences in bait disappearance were noted in field testing of the baits, indicating that decisions regarding bait formulation could be made on the basis of cost without sacrificing efficacy. Separately, testing of 17 commercially available adhesives identified five for field testing. Among these, two demonstrated strong, weather-resistant bonds between the oily surface of the artificial baits and the bait capsules as evidenced by only 13.4% and 4.5% of baits falling off suspended capsules. Importantly, neither of these adhesives impacted bait removal by snakes in field trials in comparison to baits with no adhesive.. The results represent an advance in technology development for landscape-scale suppression of brown treesnake populations.

Tpl2 kinase regulates inflammation but not tumorigenesis in mice

Tumor progression locus 2 (Tpl2, gene name MAP3K8), a mitogen-activated protein kinase, is widely expressed in immune and non-immune cells to integrate tumor necrosis factor (TNF), toll-like receptors (TLRs), and interleukin-1 (IL1) receptor signaling to regulate inflammatory response. Given its central role in inflammatory response, Tpl2 is an attractive small molecule drug target. However, the role of Tpl2 as an oncogene or tumor suppressor gene remains controversial, and its function outside immune cells is not understood. We therefore utilized a Tpl2 kinase dead (Tpl2-KD) mouse model in an 18-month aging study to further elucidate Tpl2 effects on lifespan and chronic disease. Histopathological studies revealed the incidence and severity of spontaneous tumors and non-neoplastic lesions were comparable between wild type and Tpl2-KD mice. The only finding was that male Tpl2-KD mice had higher bodyweight and an increased incidence of liver steatosis, suggesting a sex-specific role for Tpl2 in hepatic lipid metabolism. In conclusion, loss of Tpl2 kinase activity did not lead to increased tumorigenesis over aging in mice but affected likely alterations in lipid metabolism in male animals.

Spontaneous Mammary Tumor with Concurrent Cavernous Hemangioma in Albino Mice- A Rare Case Report from Mizoram, India

Background: Mouse mammary tumor is a common neoplasm of the species. Mice is an established model for human breast cancer studies and in recent times for transgenic studies. Hemangiomas are tumors of blood vascular system. Hemangioma with breast cancer is reported in human beings. Presented study describes an unusual case of concurrent mouse mammary tumor and cavernous hemangioma from Mizoram, India.Methods: A mortality of 8 out of a colony of 25 female albino mice was observed due to mammary tumors. Dead mice were thoroughly necropsied and tissue samples were collected from different parts of each tumor mass and fixed in neutral buffered formalin. The fixed tissues were processed for hematoxylin and eosin staining by routine paraffin embedding, microtomy and staining protocol. Result: Gross examination revealed variable sized tumors on abdomen of affected mice. Histopathology revealed two different forms of mammary tumor viz, acinar and solid mammary carcinoma. Most peculiar finding was remarkable presence of large, endothelial lined cavernous spaces with presence of stagnant blood. These were diagnosed as cavernous hemangioma. Perusal of available literature did not reveal any report of cavernous hemangioma occurring with mouse mammary tumor, while it is found in other species including human beings.

Blood Metabolic and Hematology Parameters and Survivorship in Mice after Application of the Rabies Challenge Virus Standard in Vaccine Potency Test

Background: Rabies virus can cause intensive and lethal infection of the central nervous system (CNS) in animals and humans. Metabolic examinations are conducted at the cerebrospinal fluid (CSF), and it has been found that many metabolic changes occur during RABV infection. However, although it is a neurotropic virus, it can cause damage to extraneural tissues - lungs, heart, kidneys and liver. This study aimed to determine differences in metabolic, endocrinology and hematologic parameters in blood of mice after application of rabies challenge with virus standard 27 strain (CVS-27).Materials, Methods &amp; Results: This study included 30 survived, and 30 dead mice that were part of the standard procedure of NIH (National Institute of Health) test in Pasteur Institute in Novi Sad. Tests were performed in the following order: two groups of mice were vaccinated in a 7 day period with different dilutions of standard vaccine and the examined vaccine. Seven days after the last vaccination, immunized animals and animals in the control group received test virus CVS-27. Blood samples were collected from a heart puncture. Differences in hematologic and biochemical parameters were determined by t-test. Due to a high number of blood parameters, we performed a joint analysis of multiple dependent variables. Higher pH value and higher concentrations of glucose, cholesterol, lactate dehydrogenase (LDH), creatine kinase (CK), albumin, urea, creatinine, α-amylase, magnesium (Mg), nonesterified fatty acids (NEFA), beta-hydroxybutyrate (BHB) and lactate were noted in dead mice. Higher granulocytes and mean platelet volume (MPV) were noted in mice which died, but also reduced lymphocytes, erythrocytes, haemoglobin, hematocrit and platelets count. Higher values of insulin, cortisol and HOMA-IR (homeostatic model assessment insulin resistance) were noted in the group of dead mice compared to the surviving one. Reduced QUICKI (quantitative insulin sensitivity check index) value was noted in mice which died compared to the surviving group. Principal component analysis (PCA) showed that components 1 and 2 explain 38.7 of variance and that these two compounds are enough for the distinction between the animals which dies and those that survived. It was found that the cortisol, insulin, HOMAIR, NEFA, aspartate aminotransferase (AST), lactic acid, LDH and granulocyte could explain the variance of the first component, which highly correlated with the first principal component. Also, pH level, glucose, creatinine, albumin and BHB showed significant importance. A positive correlation was shown between those parameters.Discussion: Mice that died during NIH test after applying CVS-27 expressed more significant stress (higher cortisol level). Disturbances of energy metabolism were noted (more significant catabolism of lipids and insulin resistance), changes of protein metabolism caused by muscle load (urea, creatinine, AST and LDH) and general disturbances of acid-base status (higher pH) and dehydration (increased albumin) were also noted in mice that died. Values of hematologic parameters showed minor influence at total variability and are a bit correlated with metabolic changes. In factor analysis, component 1 was determined from numerous parameters. Correlations between component 1 and cortisol, HOMA-IR, lactates, insulin, AST and LDH were noted. It completely determines survived and dead animals after CVS-27 during NIH test. Disturbances in blood parameters showed an analogy with previous studies of CNS. Given parameters can be very useful in clinical-pathological analysis in RABV infection.