Species of Clostridium bacteria are notable for their ability to lyse tumor cells growing in hypoxic environments. We show that an attenuated strain of Clostridium novyi (C. novyi-NT) induces a microscopically precise, tumor-localized response in a rat orthotopic brain tumor model after intratumoral injection. It is well known, however, that experimental models often do not reliably predict the responses of human patients to therapeutic agents. We therefore used naturally occurring canine tumors as a translational bridge to human trials. Canine tumors are more like those of humans because they occur in animals with heterogeneous genetic backgrounds, are of host origin, and are due to spontaneous rather than engineered mutations. We found that intratumoral injection of C. novyi-NT spores was well tolerated in companion dogs bearing spontaneous solid tumors, with the most common toxicities being the expected symptoms associated with bacterial infections. Objective responses were observed in 6 of 16 dogs (37.5%), with three complete and three partial responses. On the basis of these encouraging results, we treated a human patient who had an advanced leiomyosarcoma with an intratumoral injection of C. novyi-NT spores. This treatment reduced the tumor within and surrounding the bone. Together, these results show that C. novyi-NT can precisely eradicate neoplastic tissues and suggest that further clinical trials of this agent in selected patients are warranted.

Ford SP, Zhang L, Zhu M, Miller MM, Smith DT, Hess BW, Moss GE, Nathanielsz PW, Nijland MJ. Maternal obesity accelerates fetal pancreatic ␤-cell but not ␣-cell development in sheep: prenatal consequences. Am J Physiol Regul Integr Comp Physiol 297: R835-R843, 2009. First published July 15, 2009 doi:10.1152/ajpregu.00072.2009.-Maternal obesity affects offspring weight, body composition, and organ function, increasing diabetes and metabolic syndrome risk. We determined effects of maternal obesity and a high-energy diet on fetal pancreatic development. Sixty days prior to breeding, ewes were assigned to control [100% of National Research Council (NRC) recommendations] or obesogenic (OB; 150% NRC) diets. At 75 days gestation, OB ewes exhibited elevated insulin-to-glucose ratios at rest and during a glucose tolerance test, demonstrating insulin resistance compared with control ewes. In fetal studies, ewes ate their respective diets from 60 days before to 75 days after conception when animals were euthanized under general anesthesia. OB and control ewes increased in body weight by ϳ43% and ϳ6%, respectively, from diet initiation until necropsy. Although all organs were heavier in fetuses from OB ewes, only pancreatic weight increased as a percentage of fetal weight. Blood glucose, insulin, and cortisol were elevated in OB ewes and fetuses on day 75. Insulin-positive cells per unit pancreatic area were 50% greater in fetuses from OB ewes as a result of increased ␤-cell mitoses rather than decreased programmed cell death. Lambs of OB ewes were born earlier but weighed the same as control lambs; however, their crown-to-rump length was reduced, and their fat mass was increased. We conclude that increased systemic insulin in fetuses from OB ewes results from increased glucose exposure and/or cortisol-induced accelerated fetal ␤-cell maturation and may contribute to premature ␤-cell function loss and predisposition to obesity and metabolic disease in offspring.sheep; fetal growth; pancreatic function RECENT DATA FROM THE 1999 -2002 National Health and Nutrition Examination Survey (NHANES) show that almost 65% of the adult population in the United States is overweight, defined as having a body mass index Ͼ 25 kg/m 2 , compared with 56% observed in NHANES III, conducted between 1988 and 1994 (32a). Obesity among women of reproductive age ranges from 20 to 34% (8, 10). Furthermore, Boney et al. (8) reported that by 11 years of age, children exposed to maternal obesity were at twice the risk of developing some components of the metabolic syndrome (obesity, systolic or diastolic hypertension, high triglyceride levels, low HDL levels, glucose intolerance), which suggests that obese mothers, even in the absence of gestational diabetes, may have metabolic factors that affect fetal growth and postnatal outcomes. Maternal obesity has been associated with either intrauterine growth restriction (IUGR) or large-for-gestational-age fetuses (12, 33). Both conditions are connected to offspring exhibiting altered insulin secretion and ad...

Nonadherent marrow mononuclear cells enriched for hematopoietic progenitor cells were cultured in semisolid medium with recombinant human granulocyte-macrophage colony-stimulating factor for 9 days to form colony forming unit-granulocyte macrophage (CFU-GM) colonies. 1,25-Dihydroxyvitamin D was then gently layered over the cultures. After 2 weeks, approximately 30% of the colonies that formed were composed of cells with a unique polygonal morphology. One hundred percent of the polygonal cells in these colonies crossreacted with the monoclonal antibody 23c6, which preferentially recognizes osteoclasts. Homogenous populations of these polygonal cells formed multinucleated cells (MNC) in suspension culture, 100% of which cross-reacted with the 23c6 monoclonal antibody, and greater than 90% of the MNC contracted in response to calcitonin. Approximately 20% of these MNC formed resorption lacunae on calcified matrices. These results suggest that 1) early osteoclast precursors are derived from CFU-GM, the committed granulocyte-macrophage progenitor; 2) committed mononuclear osteoclast precursors have a distinct polygonal morphology and cross-react with monoclonal antibodies that recognize mature osteoclasts; and 3) these mononuclear precursors are capable of forming multinucleated cells which fulfill the functional criteria for osteoclasts.

Chicken infectious anemia virus (CIAV) is a resistant and ubiquitous virus of chickens causing disease in young chickens and immunosuppression in all birds. This paper reviews the current knowledge of CIAV with a focus on new findings indicating that immunosuppressive effects have not been fully appreciated, especially as they relate to the development of antigen-specific cytotoxic T cells. A more complete understanding of the immunosuppressive effects of CIAV emphasizes the need for better vaccines, especially for the broiler industry. In addition, a new model is proposed for the control of viral replication in the reproductive tract of specific-pathogen-free chickens, which may be latently infected. This model suggests that virus transcription is controlled by viral enhancer and repressor elements, which are regulated by different hormones. As a consequence, CIAV has a well-adapted relationship with its host, avoiding immune detection, ensuring passage of virus to the next generation, and eliciting limited pathology to the host.

Rift Valley fever virus (RVFV), a member of the family Bunyaviridae, causes severe to fatal disease in newborn ruminants, as well as abortions in pregnant animals; both preventable by vaccination. Availability of a challenge model is a pre-requisite for vaccine efficacy trials. Several modes of inoculation with RVFV ZH501 were tested on goats and sheep. Differences in development of infectious viremia were observed between animals inoculated with RVFV produced in mosquito C6/36 cells compared to Vero E6 cell-produced inoculum. Only C6/36-RVFV inoculation led to development of viremia in all inoculated sheep and goats. The C6/36 cell-produced RVFV appeared to be more infectious with earlier onset of viremia, especially in sheep, and may also more closely represent a field situation. Goats were somewhat more resistant to the disease development with lower and shorter infectious virus viremia, and with only some animals developing transient increase in rectal temperature in contrast to sheep. In conclusion, a challenge protocol suitable for goat and sheep vaccine efficacy studies was developed using subcutaneous inoculation of 10(7)PFU per animal with RVFV ZH501 produced in C6/36 cells.