Fur Loss Research Articles

Individual variation in life-history timing: synchronous presence, asynchronous events and phenological compensation in a wild mammal.

Many animals and plants have species-typical annual cycles, but individuals vary in their timing of life-history events. Individual variation in fur replacement (moult) timing is poorly understood in mammals due to the challenge of repeated observations and longitudinal sampling. We examined factors that influence variation in moult duration and timing among elephant seals (Mirounga angustirostris). We quantified the onset and progression of fur loss in 1178 individuals. We found that an exceptionally rapid visible moult (7 days, the shortest of any mammals or birds), and a wide range of moult start dates (spanning 6-10× the event duration) facilitated high asynchrony across individuals (only 20% of individuals in the population moulting at the same time). Some of the variation was due to reproductive state, as reproductively mature females that skipped a breeding season moulted a week earlier than reproductive females. Moreover, individual variation in timing and duration within age-sex categories far outweighed (76-80%) variation among age-sex categories. Individuals arriving at the end of the moult season spent 50% less time on the beach, which allowed them to catch up in their annual cycles and reduce population-level variance during breeding. These findings underscore the importance of individual variation in annual cycles.

A circumpolar parasite: Evidence of a cryptic undescribed species of sucking louse, Linognathus sp., collected from Arctic foxes, Vulpes lagopus, in Nunavut (Canada) and Svalbard (Norway).

The North has experienced unprecedented rates of warming over the past few decades, impacting the survival and development of insects and the pathogens that they carry. Since 2019, Arctic foxes from Canada (Nunavut) have been observed with fur loss inconsistent with natural shedding of fur. Adult lice were collected from Arctic foxes from Nunavut (n = 1) and Svalbard (n = 2; Norway) and were identified as sucking lice (suborder Anoplura). Using conventional PCR targeting the mitochondrial cytochrome c oxidase subunit 1 gene (cox1), lice from Canada and Svalbard were 100% similar (8 pooled samples from Nunavut and 3 pooled samples from Svalbard), indicating that there is potential gene flow between ectoparasites on Scandinavian and North American Arctic fox populations. The cox1 sequences of Arctic fox lice and dog sucking lice (Linognathus setosus) had significant differences (87% identity), suggesting that foxes may harbour a cryptic species that has not previously been recognised. Conventional PCR targeting the gltA gene for Bartonella bacteria amplified DNA from an unknown gammaproteobacteria from two pooled louse samples collected from Svalbard foxes. The amplified sequences were 100% identical to each other but were only 78% like Proteus mirabilis reported in GenBank (CP053614), suggesting that lice on Arctic foxes may carry unique microorganisms that have yet to be described.

Morphological, Anatomical, and Physiological Signs of Senescence in the Great Fruit-Eating Bat (Artibeus lituratus)

The longevity of bats is longer than expected based on their small body sizes and high metabolic rate. However, signs of senescence have been poorly studied for free-living bats, especially for Neotropical species, because elderly individuals are challenging to find in the wild, and long-term mark-recapture studies are lacking. Here, we present the first record of a free-living Neotropical frugivorous bat, a female Artibeus lituratus (family Phyllostomidae), with impressive signs of senescence. We compare the elderly bat with conspecific juveniles and non-elderly adults morphologically, anatomically, and physiologically. The most noticeable signs of senescence were a significant loss of fur, dark spots on the skin, excessive dental wear, and reduced ability to assimilate sugar. These traits indicate that bats' challenges as they get old can be associated with reduced thermal insulation, ability to chew, and nutrient assimilation. However, by reducing extrinsic causes of mortality, flying capability in old bats with clear signs of senescence may represent a vital survival trait associated with the long lifespan of these animals.

Leveraging Pre-Clinical Animal Model of CTCL to Explore Therapeutic Potential of a Novel STAT3 Degrader

STAT3 enhances pro-survival signaling essential for T cell expansion, but when this pathway is not appropriately regulated, aberrant STAT3 signaling contributes to T cell lymphomagenesis. Cutaneous T cell lymphoma (CTCL) is a type of mature T cell lymphoma characterized by the accumulation of malignant T cells in the skin. Our analysis, along with other recently published sequencing studies, identified upregulation of the STAT3 cytokine signaling pathway as a key driver of CTCL pathogenesis. In normal processes, STAT3 activity is transient, but dysregulation of this pathway is observed in several human malignancies, including CTCL. Aberrant STAT3 activity in malignant cells promotes transcription of target genes that promote proliferation and survival of transformed T cells. Translocations resulting in STAT3 duplications, activating mutations in the SH2 domain of STAT3 and activating mutations in kinases upstream of STAT3 all contribute to hyperactivation of this JAK/STAT signaling pathway in CTCL. STAT3 signaling in bystander cells within the tumor microenvironment may further contribute to tumor progression, although the precise role of this signaling pathway in malignant disease is context dependent. We have used conditional gene targeting to develop a fully penetrant STAT3-dependent small animal model of CTCL that recapitulates many key features of human disease. In this model, a constitutively active form of STAT3 is expressed exclusively in T lymphocytes. The autochthonous mouse model is characterized by progressive accumulation of malignant T cells in the skin and secondary lymphoid organs. Our molecular analysis of T cells from this mouse model revealed a transcriptional signature that largely mirrored the one found in malignant cells from patients. We have now taken advantage of this tractable animal model of CTCL to evaluate the therapeutic potential of a potent and selective E3 ubiquitin ligase-based novel STAT3 heterobifunctional degrader for targeting this difficult-to-treat hematologic malignancy. Weekly intravenous administration of the degrader was well tolerated and led to significant reduction of STAT3 levels in circulating and skin-resident T lymphocytes. Progression of disease in our preclinical model was notably blunted upon administration of the STAT3 degrader with dramatic reduction in T cell accumulation in the skin and lymph nodes. We will report on pre-clinical evaluation of the degrader and the impact on disease progression, T cell accumulation, activation and proliferation in vivo following weekly drug administration. Targeted protein degradation represents a novel therapeutic modality enabling direct targeting of previously undruggable oncoproteins such as STAT3. These data provide a rationale for selective STAT3 degradation as a therapeutic strategy for malignancies such as CTCL that are associated with constitutive activation of STAT3 signaling. Fig Legend:Treatment of mice exhibiting CTCL phenotypes starting at ~3mo of age. Phenotype score reflects presentation of fur loss, scaling/flaking of the skin, appearance of lesions and death. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Product Development of an Animal Shampoo from Lettuce Waste

Overexploitation of natural resources is one of the global environmental problems, so it is crucial to use waste to achieve a more sustainable world. This paper proposes the development process of a product using ingredients extracted from leaves and stems of lettuce waste. To identify and satisfy the customer needs, the Chemical Product Design procedure was followed by discussing the four stages: Needs, Ideas, Selection and Manufacture. The final product - LettPet Shampoo - consists of an Animal Shampoo that has two formats: bottles of 0.2 L and 5 L. The bioactive compounds extracted add properties to the shampoo, such as fur loss prevention, strengthening, and antioxidant properties. According to the economic analysis, 50 thousand bottles would be sold in the first year, and at the end of the 5th year, sales would be doubled. The company would obtain a positive cash flow until the 4th semester and settle the debt after three years.

Impacts of Fluoride Neurotoxicity and Mitochondrial Dysfunction on Cognition and Mental Health: A Literature Review.

This review focuses on the synthesis of current experimental and observational data regarding the effect of fluoride exposure on childhood mental health and the role of mitochondrial function as a mechanism of action. We aggregated data on the relationships between fluoride neurotoxicity, mitochondrial function, and cognitive and mental health using PubMed. Current animal and human research suggest that prenatal and perinatal fluoride exposure might have neurotoxic effects. These studies observed physical changes (fur loss and delayed reflex development in animals), intelligence loss, increased hyperactivity, and irregular moods associated with fluoride exposure. Two gaps in the literature were identified: (1) there is limited research on the mental and emotional impacts of fluoride exposure compared to research on cognitive outcomes, and (2) human studies primarily focus on prenatal and perinatal exposure, with little research conducted at other time points (e.g., adolescence). Furthermore, there is no agreed-upon mechanism for the neurotoxic effects of fluoride; however, fluoride can induce mitochondrial damage, including decreasing circulating mitochondrial DNA content, dysregulating biogenesis, and circular structure loss. Additionally, many neurodevelopmental conditions have mitochondrial underpinnings. More work is needed to elucidate the impact and timing of fluoride exposure on mental health and the role of mitochondrial function as a biological mechanism

Alvocidib Enhances Venetoclax Sensitivity in Unfavorable-Risk Multiple Myeloma Cells In Vitro and In Vivo

▪Abstract:Background: Multiple myeloma (MM) is a lymphoproliferative neoplasm involving mature plasma cells and remains incurable. New and more effective treatment options are clearly needed for higher-risk MM groups. The BCL-2-specific BH3-mimetic venetoclax (ABT-199) has been reported to be active in patients with the t(11;14) translocation, an intermediate risk group in response to immunomodulatory drugs (IMiDS) and proteasome inhibitors (PIs), prompting efforts to extend venetoclax activity to include more resistant, higher-risk MM subsets.Methods:Effects of the CDK9 inhibitor alvocidib (flavopiridol; FP) on responses to the BCL-2-specific BH3-mimetic venetoclax (ABT-199) were examined in multiple myeloma (MM) cells, including higher-risk sub-types exhibiting low BCL-2/MCL-1 expression profiles and limited sensitivity to venetoclax alone. Cell death and protein expression were evaluated by western blot, and immunofluorescence. Xenograft models, including an orthotopic murine model (NOD/SCID-γ mice) and an immune-competent model (C57BL/KaLwR1 mice) were used to characterize combination effects in vivo.Results:Alvocidib (100-200 nM; 24 h) synergistically increased ABT-199 (250-750 nM) lethality in both ABT-199-sensitive and insensitive MM cells, and these interactions were synergistic by Median Dose Effect analysis with CI values < 1.0 in both t(4;14) H929 and OPM-2 cells. Synergistic interactions or markedly enhanced lethality were also observed in multiple other intermediate or high-risk models (e.g., KMS11(t(4;14)), OPM2(t(4;14)), RPMI8226 (t(14;16) and t(8;22)), and KMS28-PE (t(4;14))). The alvocidib/venetoclax regimen was effective in various drug-resistant MM lines e.g., bortezomib, melphalan, and revlimid. Alvocidib blocked CDK9 activation and P-TEFb phosphorylation, down-regulated MCL-1, increased BCL-2/MCL-1 ratios, and up-regulated BIM. MCL-1 ectopic expression or knock-down in MM cells significantly diminished or increased ABT-199 sensitivity respectively. CDK9 shRNA knock-down triggered MCL-1 down-regulation and increased ABT-199 activity, while BIM knock-down significantly reduced alvocidib/venetoclax lethality. HS-5 cell co-culture failed to protect MM cells from the FP/ABT-199 regimen, suggesting circumvention of microenvironmental signals. The effects of venetoclax +/- alvocidib were investigated in a larger number of primary specimens (N =23). Combined treatment very significantly reduced the survival of cells compared to single agent treatment (24 h; P < 0.0001 and 0.0016). In the sub-set of specimens in which primitive progenitor cells could be identified (CD138-/CD19+/CD20+/CD127+; N=18), significant reductions in survival following combined treatment were also observed (P < 0.0074 and 0.0001). In contrast, alvocidib did not increase ABT199 lethality in normal CD34+ cord blood cells. To assess the in vivo relevance of these findings, NOD/SCID-γ mice were inoculated with fluorescently-labled bortezomib-resistant PS-R cells. Combined treatment sharply reduced tumor burden and significantly prolonged survival compared to single-agent treatment (P < 0.0001). Median survival was 79 days for animals in the combined treatment group, versus 62, 63, and 71 days for those in vehicle, venetoclax, or alvocidib groups, respectively. In addition, combined treatment was not associated with significant increases in weight loss or other forms of toxicity e.g., behavioral changes, fur loss etc. Finally, parallel studies were performed in an immune-competent model. C57BL/KaLwR1 mice were injected intravenously with murine MM 5TGM1 cells stably expressing luciferase. Combined administration suppressed tumor growth, manifested by clearly diminished intensity and significantly prolonged survival compared to single agents (P < 0.001), but did not lead to increases in toxicity e.g., weight loss. Specifically, median survival was 69 days for animals in the combined treatment group, versus 39, 49, and 53 days for those in vehicle, venetoclax, or alvocidib groups, respectively.Conclusions:These findings argue that CDK9 inhibitors e.g., FP may increase the anti-myeloma activity of ABT-199, including in unfavorable-risk MM minimally responsive to ABT-199 alone. DisclosuresOrlowski:BioTheryX: Consultancy, Membership on an entity's Board of Directors or advisory committees.

The Incidence of Dirofilaria immitis in Shelter Dogs and Mosquitoes in Austria.

To estimate the incidence of Dirofilaria immitis in Austrian shelter dogs and mosquitoes trapped in their proximity, 115 shelter dogs from fourteen animal shelters located in five different Austrian states were examined. Blood samples were screened for D. immitis using ELISA antigen-testing, PCR and microscopical examination for microfilariae. In total, 91% of the dogs originated from countries endemic for dirofilariosis. Eleven dogs (9.6%), all originating from Hungary, tested positive for D. immitis. None of the dogs examined showed microfilaremia. Eight dogs showed no or only mild clinical signs (e.g., infrequent coughing), and three dogs showed frequent coughing, dyspnea, exercise intolerance, blunt fur or weight loss. In total, 205 Mosquitoes of ten different species were caught at five different shelter sites in four different Austrian states, using CO2-baited mosquito traps set once a month (June–September 2019) for 24 h. All 205 mosquitoes tested negative for Dirofilaria spp. via PCR. The risk of endemisation of D. immitis in Austria (and other non-endemic countries in a similar situation) is very serious and its zoonotic potential should be communicated more strongly. To monitor a possible transmission of microfilariae from untreated or even untested positive dogs, e.g., in animal shelters, to mosquitoes in the near surroundings, frequent screening for Dirofilaria in mosquitoes should be used more intensively. Current knowledge on D. immitis should be integrated into daily veterinary practice and dog owners should be proactively educated, especially before traveling to endemic areas or adopting dogs from endemic countries. Animal shelters and animal welfare organizations should be provided with appropriate education and veterinary guidance regarding the testing and treatment of dogs imported from high-risk areas.

Fur loss syndrome and lice infestations observed on Arctic foxes in central Nunavut, Canada

As temperatures in the circumpolar north continue to warm, shifts in species distribution and the breakdown of environmental barriers for arthropods may impact the diversity and distribution of ectoparasites in Arctic ecosystems. In May 2019, fur loss over the neck and shoulders was observed on Arctic foxes in a terrestrial Arctic ecosystem (Karrak Lake) in central Nunavut, Canada. This was inconsistent with normal patterns of shedding winter fur and had not been observed on Arctic foxes in this population over the previous 19 years of live-trapping. Operculated eggs attached to hair shafts were collected from one affected fox. Conventional PCR using universal louse primers targeting conserved regions of mitochondrial 12S and 16S rDNA confirmed that the eggs belonged to the order Phthiraptera. Sequencing results were inconclusive at the species level. Further investigation revealed a single unpublished report of an Arctic fox with similar fur loss trapped on mainland Nunavut, in 1997. Adult lice collected from this fox were identified as sucking lice (potentially from the genus Linognathus). Our findings emphasize the need for further monitoring and have significant implications for trappers and wildlife management, as infestations negatively impact the pelt quality of these important furbearers.

Standardized experimental model for cement dust exposure; tissue heavy metal bioaccumulation and pulmonary pathological changes in rats.

A controlled experimental model of exposure to aerosols particularly for cement dust was recently invented in a study from the laboratory that found high serum levels of heavy metals, decrease gastrointestinal motility, and altered hematological variables in cement dust exposed rats. However, reproducibility was not considered. This work aims at standardizing the model and investigating preliminary toxicological indicators. Thirty male rats used in this study were divided into 3 groups (n = 10). Group 1; control, while groups 2 and 3 were exposed to cement dust for 14 days and 28 days respectively. We assessed clinical signs of toxicity, tissue heavy metal concentration, histopathological, and body weight (BW) changes. We observed poor movement coordination, abnormal posture, cephalic fur loss. Evidence of ischemia and fibrotic pneumoconiosis were grossly observed in the lungs of the exposed groups. There was a significant increase in tissue level of heavy metals with pulmonary and gastric heavy metal content showing a trendy relationship during the period of the exposure as the value of Lead, Chromium, Cadmium, Iron, Calcium, and Nickel increased by nearly similar percentages in both tissues. Organs weights increased; the 14-day exposed (198 ± 31; 168 ± 22) and 28-day exposed (198 ± 22; 187 ± 26) groups had significantly reduced body weight at the first and second weeks of exposure compared to the control group (265 ± 26; 357 ± 40) respectively. Exposure to cement dust induced low bone density in the exposed rats (p < 0.05). Histopathological alterations include necrosis, inflammatory cellular infiltration, and alveolar hyperplasia suggestive of the proliferative response of pulmonary tissue to the dust. The operation of the standardized apparatus mimics a typical occupational exposure and the findings show that cement dust induces systemic toxicity via respiratory perturbation and body/organ weight discordance mediated by heavy metal bioaccumulation.

Free Rather Than Total Iron Content Is Critically Linked to the Fur Physiology in Shewanella oneidensis.

Ferric uptake regulator (Fur) is a transcriptional regulator playing a central role in iron homeostasis of many bacteria, and Fur inactivation commonly results in pleiotropic phenotypes. In Shewanella oneidensis, a representative of dissimilatory metal-reducing γ-proteobacteria capable of respiring a variety of chemicals as electron acceptors (EAs), Fur loss substantially impairs respiration. However, to date the mechanism underlying the physiological phenomenon remains obscure. This investigation reveals that Fur loss compromises activity of iron proteins requiring biosynthetic processes for their iron cofactors, heme in particular. We then show that S. oneidensis Fur is critical for maintaining heme homeostasis by affecting both its biosynthesis and decomposition of the molecule. Intriguingly, the abundance of iron-containing proteins controlled by H2O2-responding regulator OxyR increases in the fur mutant because the Fur loss activates OxyR. By comparing suppression of membrane-impermeable, membrane-permeable, and intracellular-only iron chelators on heme deficiency and elevated H2O2 resistance, our data suggest that the elevation of the free iron content by the Fur loss is likely to be the predominant factor for the Fur physiology. Overall, these results provide circumstantial evidence that Fur inactivation disturbs bacterial iron homeostasis by altering transcription of its regulon members, through which many physiological processes, such as respiration and oxidative stress response, are transformed.

Abstract 5049: Development of huPBMC-NCG mice to study xenogenic graft versus host disease (GVHD)

Abstract Graft versus host disease (GVHD) is a severe complication of allogeneic stem cell transplantation for leukemia treatment. NCG mice lacking functional murine T, B, NK cells as well as decreasing functional macrophage and dendritic cells could be used as recipients of human peripheral blood mononuclear cells (huPBMC) for the studies of human GVHD. In this study, huPBMC-NCG mice were developed by the engraftment of huPBMC into unconditioned or irradiated NCG mice. The engraftment kinetics, irradiation effect, and preclinical treatment were explored in these mice. After huPBMC engraftment, NCG mice developed GVHD symptoms, including body weight loss, fur loss, reduced activity, hunched posture and severe ruffling, and eventually led to mortality. We also observed human T cell trafficking to target organs including gut, heart, liver, skin, and lung by flow cytometry and immunohistochemistry. Interestingly, irradiation could significantly accelerate the onset of GVHD following huPBMC engraftment. Meanwhile, irradiated huPBMC-NCG mice presented less variability in the kinetics of GVHD and the frequency of this disease in comparison with unconditioned counterparts, thus providing a more stable model for human GVHD studies. For preclinical studies, we found immunomodulatory drugs including Abatacept (CTLA4-Ig) targeting APC cells or anti-Ly6G antibody targeting neutrophils could alleviate GVHD-associated symptoms along with improving overall survival of huPBMC-NCG mice. Taken together, this study established stable huPBMC-NCG mice to mimic the onset and development of human GVHD, which was helpful for the development of therapeutic intervention. Citation Format: Cunxiang Ju, Hongyan Sun, Dingyu Wang, Shiying Guo, Mingkun Zhang, Hainan Wu, Shuai Li, Chao Ju, Jing Zhao, Xiang Gao. Development of huPBMC-NCG mice to study xenogenic graft versus host disease (GVHD) [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5049.

Abstract 248: Prospective Study of Ventricular Function and Survival in Acute Phase of Chagas Disease: An Animal Experimental Model

Background: Acute Cardiomyopathy secondary to Chagas disease is often subclinical when related to vector transmission. However, cases related to non-vector transmission as in patients submitted to cardiac transplant may have fatal outcomes. This study aimed to assess survival during the acute phase of Chagas disease and its relation to the ventricular function in an animal experimental model. Methods: Female Syrian hamsters (n=45) were separated in two groups: control group (CG):15 animals injected with saline solution; and infected group (IG): 30 animals inoculated with 3,5x10 4 trypomastigote forms of Trypanosoma cruzi,Y strain . Both groups were monitored daily and submitted to echocardiography with equipment dedicated to small animals (Vevo® 2100) in two different moments: baseline (before infection) and 15 days post infection. Left ventricular ejection fraction (LVEF) and global longitudinal myocardial strain (GLS) of left ventricle were measured. The IG was divided into animals with and without clinical sign (CS+) of disease: weight or fur loss, mucous wounds and lethargy. ANOVA for mixed models was used to compare the ventricular function parameters among groups over time. Survival analysis was studied using Kaplan-Meier curves and logrank test. Results: Total time follow up was 60 days. LVEF in IG was significantly reduced through time (53.80 ± 4.95 to 43.55 ± 12.10%) compared to CG (57.86 ± 7.52 to 59.73 ± 5.87%) (p=0.002). There was also a significant reduction of GLS (-18.97 ± 3.94 to -12.44 ± 4.79%) in the IG compared to CG (-19.58 ± 4.03 to -19.67 ± 4.04%) (p=0.012). Twelve animals from IG died (40.00%,12 out of 30) compared to one from CG (6.66%, 1 out of 15). Eleven out of the 12 dead animals from IG, presented, before, clinical signs (CS+). Survival was significant reduced in the IG compared to CG over time (p=0.02) (Figure 1). Conclusion: Reduced survival during the acute phase of experimental model of Chagas disease is related to the significant reduction of left ventricular function. The mortality rate in the IG is higher in the group which presents CS+.

The Role of Scarpa’s Fascia in the Human Body Wall

Humans have lost a cutaneous muscle layer called the panniculus carnosus that is found in a large survey of mammals and many non‐human primates. Humans have a tough fascial layer called Scarpa’s fascia in the same position as panniculus carnosus, which some researchers suggest is the remnant of this cutaneous muscle layer. In other mammals, panniculus carnosus shakes the skin, possibly to dislodge ectoparasites. However, in humans, it is possible that a tough fascia is more adaptive for passively maintaining the human orthograde posture than an isotonic muscle layer, and the loss of fur on the skin may require less movement to combat parasites. Interestingly, panniculus carnosus is generally associated with a fatty fascial layer termed panniculus adiposus. Traditionally, Camper’s fascia, a fatty layer just deep to the skin and superficial to Scarpa’s fascia has been assumed to be the representative of panniculus adiposus in human. However, our dissections demonstrate an undescribed fatty layer deep to Scarpa’s fascia. Based on comparative anatomy of the anterior abdominal wall across several groups of mammals, and the continuity of this fatty layer into the human perineum from the abdomen, we suggest that this undescribed deeper layer may be the evolutionarily conserved panniculus adiposus and not Camper’s fascia.Support or Funding InformationFunded by Midwestern University, University of Central Florida, and the American Association for Anatomy

The therapeutic effects of edaravone on collagen-induced arthritis in rats.

Current research suggests that synovial phagocytic cells remove excessive amounts of free oxygen radicals (reactive oxygen species [ROS]), thereby preventing damage to synovial tissues. Moreover, ROS may affect the expression of growth arrest and DNA damage inducible α (GADD45A), thus further promoting the activation of synovial fibroblasts. Male adult rats were assessed for progression of collagen-induced arthritis (CIA) using a macroscopic arthritis scoring system of the hind paws and by measuring the changes in the rat's body weight, and activity level before and after diagnosis of CIA. Rats were intraperitoneally injected twice daily with edaravone at doses of 3, 6, and 9 mL/kg. Samples were taken at 2, 4, and 6 weeks, respectively. Edaravone was found to significantly reduce macroscopic arthritis and microscopic pathology scores in CIA rats. The concentration of endothelial nitric oxide synthase-6, glutathione, and heme oxygenase-1 in the serum of rats decreased, as was the production of ROS around the synovium and inflammatory factors. Moreover, ROS-1 increased the expression of the nuclear factor-κB (NF-κB) p65 protein by altering the expression level of GADD45A, causing aggravation of tissue damage. Edaravone also significantly improved the physiological condition of CIA rats, including appetite, weight changes, and loss of fur, as well as limb mobility. We believe that edaravone acts to reduce the expression of NF-ĸB p65 by clearing ROS, which causes reduced expression of GADD45A, and subsequently reduces the level of apoptosis and inflammatory response proteins, thereby reducing the symptoms of CIA. We, therefore, propose that edaravone is an effective option for clinical treatment of rheumatic arthritis.

Analysis Activity of Elephantopus scaber Leaves Extract Against Quantitative Changes of Lymphocytes Cells in BALB/c Mice After Induction of DMBA and Estrogen

The carcinogenic compounds such as DMBA known for having the ability to promote the DNA damage that affects the growth regulator genes. Therefore, uncontrolled cell growth will occur and change the immune system. Elephantopus scaber extract is considered as an alternative way of handling breast cancer since it capable to affect the work of the immune system. Hence, this study aims to determine the effect of E. scaber extract on quantitative changes of CD4 + , CD8 + T cells and proinflammatory cytokines CD4 + IFN-γ + and CD4 + TNF-α + after induction of DMBA and estrogen. In vivo experiments were carried out by using 5-6 weeks old of female mice BALB/c strain. The extract of E. scaber leaf powder was obtained from Material Medica Batu, Malang. The experimental animals were divided into five groups such as negative control group (K-), positive control group (K+), mice treated with E. scaber extract for a week (group 1), mice treated with E. scaber extract for 2 weeks (group 2), and mice treated with E. scaber extract for 3 weeks (group 3). Induction of DMBA and estrogen was conducted by injection of DMBA (7.12 dimetylbenz (α) anthracene) 0.56 mg.kgBW -1 and estradiol hormone 0.0504 mg.kgBW -1 . Mice were treated with E. scaber extract with a dose of 50 mg.kgBW -1 . Lymphocytes were isolated from spleen and observed the CD4 + , CD8 + T cells and proinflammatory cytokines CD4 + IFN-γ + and CD4 + TNF-α + T cells through flow cytometry analysis. One-way ANOVA (p <0.05) and SPSS were used to analyze this data. The results showed that the induction of DMBA and estradiol hormone affected mice fur loss. It also decreased the immune system by lowering the number of CD4 and CD8 T cells. However, E. scaber extracts increased the number of pro-inflammatory cytokines TNF-α. Furthermore, the DMBA and estradiol induction also promoted the number of CD4 + IFN-γ + in the first week, but decreased the number of CD4 + IFN-γ + at weeks 2 and 3, then increase the number of CD4 + IFN-γ + . So, the treatment of E. scaber extracts demonstrated the ability to restore the homeostasis of the immune system in the first and second weeks. Keywords: CD4 + , CD8 + , DMBA, estrogen, E.scaber, IFN-γ + , TNF-α +

Recovery of southern bent-winged bats (Miniopterus orianae bassanii) after PIT-tagging and the use of surgical adhesive

Surgical adhesive is widely used to prevent shedding of injected PIT-tags, but the effect of this adhesive on individuals has not been documented. We recaptured 52 southern bent-winged bats up to 741 days after PIT-tagging. All recaptured individuals were in good body condition, with no signs of infection or skin irritation; however, temporary fur loss occurred at the injection site. We advocate that researchers consider the impact this may have on study species, particularly when PIT-tagging small mammals during cooler months.

Abstract 2342: Experimental modeling of acute- and chronic-GvHD by xenotransplanting human donor PBMCs or cord blood CD34+ cells (HSC) into NSG mice

Abstract Graft vs. host diseases (GvHD) accounts for 15-30% of deaths following allogenic hematopoietic stem cell transplantation (allo-HSCT) for treatment of malignant diseases. Acute GvHD (aGvHD) typically involves skin, gastrointestinal and hepatic inflammation, and occurs within 100 days of transplantation; Chronic GvHD (cGvHD) involves multiple organs and occurs beyond 100 days. aGvHD is largely due to the rapid activation of donor T cells (Th1, CD8+ biased), causing tissue damage and often leading to mortality; in contrast, cGvHD is Th2 biased, typically display autoimmune-like syndrome, involving both T- and B-cell. Currently, GvHD models are mostly allo-transplantations between mice, which are criticized for poor physiological relevance. Here, we present Xenotransplantation of human donor into NSG mice, which will address some of the limitation of current models. Human peripheral blood mononuclear cells (PBMC) derived from normal donors were transplanted into NSG or NSG-like strains for modeling aGvHD. Cord blood derived HSCs (hCD34+) NSG (Jackson) were transplanted for modeling cGvHD. Hosts were monitored twice weekly, including clinical observations (e.g. animal postures, activity, fur texture, and skin integrity), body weight changes, gross pathology and histopathology upon termination, along with human-immunological phenotyping of peripheral blood, spleen, lung, liver and skin by flow cytometry, histopathology and/or immunohistochemistry. In addition, GvHD novel biomarkers Elafin, IL-18, REG3α and ST2, measured by ELISA (MBL International) at baseline and post cells engraftment. NSG mice engrafted with human PBMC, or purified T-cells, from normal donors rapidly developed typical symptoms of aGvHD, as early as 4 weeks post transplantation, including severe body loss, reduced activity, hunched posture, loss of fur, severe ruffling and overall poor grooming. Significant engraftment of human CD45+ cells were detected up to 6 weeks post-engraftment dominated with CD3+ human T-cells of single positive CD4+ or CD8+ T-cells. The corresponding kinetics of clinical symptoms in parallel with the degree of engraftment of T cells suggests xenografting were responsible for aGvHD. As for the cGvHD, NSG-mice engrafted with CD34+ cells derived from cord blood with cGvHD high- risk HLA haplotypes C*0602 and C*0401. Manifestation of cGvHD was observed 18 weeks to 39 weeks post-engraftment; symptoms were similar to aGvHD, but also include facial/full body alopecia, and scaly skin. Interestingly, in correlation of this, human engraftment of CD45+, particularly T-cells including CD4+ and their CD30+ subsets, are kinetically increased in blood and spleen with similar timeline, suggesting their roles in the observed cGvHD. The xenograft murine model using adult human PBMC and cord blood derived CD34+ HSCs could be alternative experimental systems to model human aGvHD and cGvHD for investigating disease mechanisms and evaluating treatment strategy. Citation Format: Ann E. Lin, Annie X. An, Mingfa Zang, Derron Yu, Eunmi Hwang, Israel Romero, Linda Quirino, Marybeth George, Pirouz Daftarian, Wenqing Yang, Henry Li. Experimental modeling of acute- and chronic-GvHD by xenotransplanting human donor PBMCs or cord blood CD34+ cells (HSC) into NSG mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2342.

Biodistribution and Systemic Effects in Mice Following Intravenous Administration of Cadmium Telluride Quantum Dot Nanoparticles.

Quantum dots (QDs) are engineered nanoparticles (NPs) of semiconductor structure that possess unique optical and electronic properties and are widely used in biomedical applications; however, their risks are not entirely understood. This study investigated the tissue distribution and toxic effects of cadmium telluride quantum dots (CdTe-QDs) in male BALB/c mice for up to 1 week after single-dose intravenous injections. CdTe-QDs were detected in the blood, lung, heart, liver, spleen, kidney, testis and brain. Most CdTe-QDs accumulated in the liver, followed by the spleen and kidney. At high doses, exposure to CdTe-QDs resulted in mild dehydration, lethargy, ruffled fur, hunched posture, and body weight loss. Histological analysis of the tissues, upon highest dose exposures, revealed hepatic hemorrhage and necrotic areas in the spleen. The sera of mice treated with high doses of CdTe-QDs showed significant increases in alanine aminotransferase (ALT), aspartate aminotransferase (AST), and total bilirubin levels, as well as a reduction in albumin. CdTe-QD exposure also led to a reduced number of platelets and elevated total white blood cell counts, including monocytes and neutrophils, serum amyloid A, and several pro-inflammatory cytokines. These results demonstrated that the liver is the main target of CdTe-QDs and that exposure to CdTe-QDs leads to hepatic and splenic injury, as well as systemic effects, in mice. By contrast, cadmium chloride (CdCl2), at an equivalent concentration of cadmium, appeared to have a different pharmacokinetic pattern from that of CdTe-QDs, having minimal effects on the aforementioned parameters, suggesting that cadmium alone cannot fully explain the toxicity of CdTe-QDs.

Preclinical safety of ginsenoside compound K: Acute, and 26-week oral toxicity studies in mice and rats

Ginsenoside compound K (CK) is a hydrolysate of ginsenosides in the soil bacteria. This study evaluated the toxicity of CK as acute and the 26-week repeated-dose. The results of acute toxicity show that CK administered orally to rats and mice did not cause mortality or toxicity at the maximum dosage of 8 g/kg and 10 g/kg, respectively. In the toxicity study for 26-week, rats were administered with CK at doses of 13, 40, or 120 mg/kg, and were observed for 26 weeks and recovery periods of four weeks. Under the conditions, asthenia, hypoactivity, loss of fur and body weight reduction were transiently noticed in males of 120 mg/kg group. Hepatotoxicity and nephrotoxicity also were evident including the elevation of liver and kidney relative weight, along with focal liver necrosis as well as the increase in plasma enzymes (ALT and ALP) in male rats receiving CK (120 mg/kg), but this toxicity might be reversible. For 13 and 40 mg/kg CK groups, there was no significant variation in food habits, clinical signs, urine analysis, body weight, biochemical and hematological values, organ coefficient and histopathology examination. The NOAEL for male and female rats were observed to be 40 and 120 mg/kg, respectively.