Effect of diet and animal care/housing protocols on body weight, survival, tumor incidences, and nephropathy severity of F344 rats in chronic studies

Diet is an important environmental factor affecting body weight, survival, and age-related diseases of rodents. The NIH-07 open formula diet was the diet used in the National Toxicology Program's (NTPs) rodent carcinogenicity studies from 1980 to 1994. In 1994 the NTP began using a new diet designated the NTP-2000 diet. This paper compares body weight, survival, tumor incidence, and nephropathy severity in untreated control groups of Fischer 344 (F344) rats fed the NTP-2000 or NIH-07 diets, using data from 22 separate 2-year feed and inhalation studies. The feed studies were conducted in 3 different facilities, and all the inhalation studies were conducted in a single facility. During feed studies, rats were group housed in polycarbonate cages and fed diets in powder (mash) form, while in inhalation studies, rats were housed individually in wire mesh cages, and fed diets in pelleted form. Survival was significantly (p<0.05) higher in groups fed NTP-2000 diet compared to the corresponding groups fed NIH-07 diet, irrespective of sex or housing conditions. Use of the NTP-2000 diet was also associated with a decreased incidence of pituitary gland tumors in both sexes and decreased incidences of adrenal pheochromocytoma and preputial gland tumors in males. The incidence and severity of nephropathy was also decreased in animals receiving the NTP-2000 diet, especially males. The decreased nephropathy severity and the decreased incidence of pituitary gland tumors are likely the major factors contributing to the improved survival of rats receiving the NTP-2000 diet relative to those given the NIH-07 diet. These data also support earlier findings that decreased incidences of adrenal pheochromocytoma are associated with reduced nephropathy severity in male F344 rats. Throughout the two-year study female rats receiving the NTP-2000 diet were significantly (p<0.05) lighter than those receiving the NIH-07 diet. However, it is uncertain if this difference can be attributed to the NTP-2000 diet, since implementation of this diet by the NTP approximately coincided with changes in the F344 rat production colony that resulted in somewhat lighter animals being provided to the NTP. Controls from inhalation studies and feed studies differed significantly (p<0.01) in the incidence of a variety of tumors, irrespective of diet. This suggests that differences in animal care and housing protocols may impact tumor incidence in F344 rats, most notably pituitary gland and testis tumors.     

In vitro effect of monoclonal anti-idiotype antibodies (anti-M104E) on MOPC 104E myeloma cells

We investigated the effect of three monoclonal anti-idiotype antibodies (anti-M104E) on various functions of MOPC 104E myeloma cells in vitro. The antibodies used were N-20-2 [immunoglobulin M (IgM), BALB/c], SJL18-1 [IgM, BALB/c X SJL F1], and CD3-2 [immunoglobulin G1 (IgG1), BALB/c X A/J F1]. The two IgM antibodies were very efficient in blocking surface M104E IgM as shown by rosette inhibition, whereas the IgG1 isotype was not very effective. The reexpression of surface M104E IgM was different from antibody to antibody. The secretion of M104E IgM by MOPC 104E cells was partially blocked by the two IgM antibodies, but the IgG1 antibody had no effect. All three anti-idiotype antibodies inhibited the stem cell renewal activity of MOPC 104E cells assayed by colony formation assay. On the other hand, in suspension culture, the two IgM antibodies inhibited the growth of MOPC 104E cells in the absence of complement or effector cells of antibody-dependent cellular cytotoxicity, but IgG1 antibody had no effect. The starting tumor inoculum size was critical in the observations of the effects seen on both the growth and the colony-forming activity of MOPC 104E cells. The results of this study show the functional differences between various monoclonal anti-idiotype antibodies and also indicate that some anti-idiotype antibodies can inhibit the growth of MOPC 104E myeloma cells directly without any help of complement or effector cells of antibody-dependent cellular cytotoxicity.     

The identity of the unconditioned stimulus to the central nervous system is interferon-β

The specific mechanism of interaction between the central nervous system and immune system was examined using conditioned augmentation of natural killer (NK) cell activity. This study focused on the role of interferon-β (IFN-β) as the unconditioned stimulus (US). IFN-β was found to be the signal responsible for the bidirectional communication which links the central nervous system with the immune system. This was substantiated by injection of small quantities of IFN-β directly into the cisterna magna, which activated the effector pathway from the central nervous system to the immune system. More importantly, we found that when the conditioned stimulus (CS) was paired with an injection of IFN-β into the cisterna magna, the conditioned animals were able to raise their natural killer cell activity in response to subsequent exposure to the conditioned stimulus. These studies show the unconditioned response must be the response of the central nervous system (CNS) to the unconditioned stimulus and not the direct effect of the substance injected into the periphery.     

Effect of reserpine on retention of the conditioned NK cell response

The effect of reserpine and 6-hydroxydopamine on the learned conditioned natural killer (NK) cell response was investigated in mice. Reserpine given at 2.5 mg/kg, 24 hr prior to reexposure to camphor-conditioned stimulus on days 6 and 8 blocked the recall of conditioned NK cell response to a significant extent. In other words, the NK cell activity of conditioned mice, treated with reserpine and reexposed to the conditioned stimulus, was similar to the nonconditioned (NC) group. A conditioned increase in NK cell response was still evident in mice treated with 6-OHDA.     

Regulation of mopc 104e by T cells and growth factors induced by C. Parvum stimulation

Corynebacterium parvum, known for its ability to retard the growth of experimental neoplasms, was examined for its effect on the growth of murine plasmacytoma MOPC 104E in vivo and in vitro. Immunostimulation with C. parvum² resulted in the accelerated growth of MOPC 104E in experimental animals, as measured by the tumor IgM production. The acceleration of plasmacytoma growth was seen in all cases where C. parvum was given before or at the time of tumor transplantation. Increased proliferation of MOPC 104E was also observed when MOPC 104E was co-cultured in the presence of spleen cells from C. parvum stimulated mice as compared to the normal spleen cells. Removal of T cells by in vivo anti-thymocyte serum treatment, followed by anti-Thy 1.2 and complement in vitro, resulted in the partial loss of stimulatory activity. Furthermore, the stimulatory activity was shown to be associated with soluble mediators, which were generated by splenic adherent cells and T cells, and were, at least in part, responsible for the growth of plasmacytoma. Normal spleen cells did not generate a significant amount of soluble factor, but were able to augment MOPC 104E growth in co-culture at high spleen to tumor cell ratio.     

Modulation of immune response. A possible role for murine salivary epidermal and nerve growth factors

Both epidermal growth factor (EGF) and nerve growth factor (NGF) were found to moderate the reactivity of mouse spleen cells in an in vitro model of immune response. Blastogenic response was suppressed by EGF and potentiated by NGF. Both proteins are present in mouse, but not human, saliva in quantities equal to or greater than those affecting blastogenic response in our assay. Immune response occurring in the mouse periodontium may be affected by the presence of these factors in the saliva.     

Identification and synthesis of potential impurities of rabeprazole sodium

Rabeprazole sodium (1, Achiphex) is a gastric proton pump inhibitor. It causes dose-dependent inhibition of acid secretion and is useful as an anti-ulcer agent. In the process for the preparation of 1, two potential unknown impurities were identified in HPLC at levels ranging from 0.05-0.8%. Based on mass spectral data vide LC-MS, the two impurities were characterized as 2-{[(4-chloro-3-methyl-2-pyridinyl) methyl] sulfinyl}-1H-bezimidazole (2, chloro analogue of rabeprazole) and 2-[{(4-methoxy-3-methyl-2-pyridinyl)methyl}sulfinyl]-1H-benzimidazole (3, methoxy analogue of rabeprazole). The structures were unambiguously established by independently synthesizing them and co-injecting in HPLC. To our knowledge, the compounds 2 and 3 have not been reported as process impurities elsewhere.     

Autoimmune disease antigens1

Abstract. The first step towards understanding the cellular interaction which results in autoimmune disease is to determine what triggers the recognition between a specific autoimmune antigen determinant and the cellular receptor. In this review, we have focused on the antigen inducing experimental allergic encephalitis (EAE) because the antigen has been characterized and a relatively large body of information on its biological activities has been accumulated. Clearly, a specific allergic encephalitis-producing determinant is present and is represented on a relatively small portion of the molecule. The determinant induces a wide variety of biological reactivities, some of which are classed as cellular mediated. An attempt is made to dissect activities such as blast transformation (BT), migration inhibitory factor (MIF), in vivo delayed type hypersensitivity reaction (DTH) and EAE and to relate them to the structural requirements which the determinants possess. The complexities which arise indicate that subpopulations of cells with different receptor activities may respond selectively and that recognition of the receptor is produced by an EAE determinant consisting of three amino acids in a specific linear sequence. Furthermore, under experimental circumstances the EAE activity can be dissociated from the other activities (BT, MIF, DTH), indicating that while these tests are used generally to follow various human autoimmune disease activities, they may represent the reaction of a broad spectrum of cells.