Bone Marrow Transplantation Restores Follicular Maturation and Steroid Hormones Production in a Mouse Model for Primary Ovarian Failure

Recent studies suggest that bone marrow stem cells (BMSCs) are promising grafts to treat a variety of diseases, including reproductive dysfunction. Primary ovarian failure is characterized by amenorrhea and infertility in a normal karyotype female, with an elevated serum level of follicle-stimulating hormone (FSH) and a decrease level of estrogen caused by a mutation in FSH receptor (FSHR) gene. Currently, there is no effective treatment for this condition. The phenotype of FSHR (−/−) mouse, FORKO (follitropin receptor knockout), is a suitable model to study ovarian failure in humans. Female FORKO mice have elevated FSH, decreased estrogen levels, are sterile because of the absence of folliculogenesis, and display thin uteri and small nonfunctional ovaries. In this study, we determined the effects of BMSC transplantation on reproductive physiology in this animal model. Twenty four hours post BMSC transplantation, treated animals showed detectable estroidogeneic changes in daily vaginal smear. Significant increase in total body weight and reproductive organs was observed in treated animals. Hemotoxylin and eosin (H&E) evaluation of the ovaries demonstrated significant increase in both the maturation and the total number of the follicles in treated animals. The FSH dropped to 40–50% and estrogen increased 4–5.5 times in the serum of treated animals compared to controls. The FSHR mRNA was detected in the ovaries of treated animals. Our results show that intravenously injected BMSCs were able to reach the ovaries of FORKO mice, differentiate and express FHSR gene, make FSHR responsive to FSH, resume estrogen hormone production, and restore folliculogenesis

Canine Leishmaniasis Caused by <i>Leishmania Leishmania Infantum</i> in Two Labrador Retrievers

Canine leishmaniasis, a generally fatal parasitic disease, was diagnosed in 2 dogs with a medical history of foreign travel, lymphadenopathy, emaciation, anorexia, intermittent fever, and cutaneous lesions. Clinically, hyperproteinemia, proteinuria, azotemia, and glomerulopathy were evident. Isolation of Leishmania species was done using Schneider's Drosophila medium. Syrian hamsters were used for infectivity studies. Clear taxonomic identification was done biochemically by isoenzyme analysis and comparison of zymogram banding patterns with 6 World Health Organization reference strains. Based on the geographic origin of affected dogs, clinicopathologic presentation, visceralization with hepatosplenomegaly in hamsters, and isoenzyme analysis, a diagnosis of Leishmania leishmania infantum was made. This study, representing the first taxonomic identification of an isolate from canine leishmaniasis, demonstrates the Zoonotic and epidemiologic implications of this disease. </jats:p

Abstract 1278: Obesity enhances benzo(a)pyrene-induced colon tumorigenesis in a PIRC rat model of colon cancer..

Abstract Colorectal cancer is a contributing factor for significant mortalities in the United States, with 50,000 deaths per year and around 140,000 new cases expected to be diagnosed in this year. Epidemiological studies have pointed out that obese people have a higher risk for colon cancer. It is well known that overweight or obesity is induced by excess energy intake from dietary fat. Additionally, consumption of well-done red meat and saturated fats, rich in food-borne environmental toxicants such as polycyclic aromatic hydrocarbons (PAHs) has also been implicated as one of the causative factors for sporadic colon cancer. Thus, on one hand, diet-induced obesity and on the other hand exposures to environmental carcinogens contribute to the development of colon cancer. Therefore, the objective of this study was to investigate whether the colon polyp burden was accelerated by diet-induced obesity when exposed to environmental carcinogens.To test this concept, we have employed an adult male transgenic rat model, the Polyposis In the Rat Colon (PIRC) kindred type. This rat is a mutagen-induced nonsense allele of the rat Apc gene on an inbred F344/NTac (F344) genetic background. This rat model is advantageous because of its longer life span and lack of the tumor suppressor gene Apc. We have explored whether tumor burden in PIRC male rat (PIRC-M Heterozygous F344/NTac-Apcam1137) was influenced by the ingestion of different types of fat containing benzo(a)pyrene [B(a)P], a prototypical PAH compound. Treatment consisted of 25 and 50 μg B(a)P/kg body wt., dissolved in tricaprylin, administered to 7-week-old male PIRC rat daily via oral gavage for 60 days. One group of rats received the AIN-76A control diet and the other group, the Western diet throughout the duration of this study. At the end of exposure, rats were sacrificed; colons were retrieved and preserved in 10% formalin for observation of gross pathological changes. An increased prevalence of adenomas in colon of rats that were maintained on Western diet compared to AIN-76A diet and controls (P &amp;lt; 0.05) was noticed. Interestingly, we have also observed adenomas with high grade dysplasia in B(a)P + Western diet group and these incidences were of frequent occurrence at 50 μg/kg compared to 25 μg/kg B(a)P dose group. On the other hand, the B(a)P alone, and AIN-76A diet groups did not show significant differences in the numbers of adenomas and invasive tumors in colon. Also, B(a)P treatment-related changes were seen in body weight and food consumption of rats administered with B(a)P, with the changes more pronounced in the body weight gain of Western diet group compared to the AIN-76A diet group and controls (p &amp;lt; 0.005). In summary, our studies established that Western diet potentiates the development of colon tumors caused by B(a)P in the PIRC rat. Citation Format: Kelly L. Harris, Mohammad S. Niaz, Awadh A. Binhazim, Mary K. Washington, Samuel E. Adunyah, Aramandla Ramesh. Obesity enhances benzo(a)pyrene-induced colon tumorigenesis in a PIRC rat model of colon cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1278. doi:10.1158/1538-7445.AM2013-1278</jats:p

Amplified DNA from different tissues of animals transplanted by (+/+) or (−/−) donors.

<p>Amplified DNA from different tissues of animals transplanted by (+/+) or (−/−) donors.</p

mRNA expression of FSHR gene in the ovaries of treated and control animals and in non-reproductive organs (liver) at two different time points (1 and 12 weeks).

<p>As shown, FSHR mRNA is only expressed in the ovaries of BM treated animals and there is no mRNA expression in the ovary of untreated and in non-reproductive organs of treated group.</p

Changes of total body weight(A), ovaries(B), uterus(C), vagina and cervix(D), and serum level of FSH(E) and estrogen(F) in treated (Tr) Vs control (Ct) animals at different time points of experiment.

<p>For B, C and D organs weight considered as % of total body weight. Both treated and control group had increase in total body weight but BMT group showed significantly more increase than control group (A, P<0.03). As indicated, reproductive organs which are highly modulated by estrogen, showed remarkable increase in weight at all time points of the experiment except for the first week (for B, C, and D, a P value of less than 0.04 obtained). Bone marrow transplanted animals compare to untreated controls showed 40–50% decrease in serum FSH level (E, P<0.03) and 4–5.5 folds increase in serum estrogen (F, P<0.004) at all time points of experiment.</p

Development of the ovary in BMT animal (A) compared to untreated control animal (B).

<p>Both the total number of follicles and the number of antral follicles are significantly higher in BMT compare to control group. Histological evaluation showed on average 28±4 follicles/ovary in treated group with 8±2 follicles at the antral stage compared to only 6±2 with no follicles at antral stage in untreated control mice. Photos have been taken at the same magnification.</p