Mukunda B. Ray

Efficacy of a Transforming Growth Factor β2 Containing Nutritional Support Formula in a Murine Model of Inflammatory Bowel Disease

Objective: Dietary, environmental and genetic events may influence host susceptibility to inflammatory bowel diseases (IBD). Transforming growth factor β2 (TGF-β2), a multifunctional polypeptide (cytokine) present in human and bovine milk, plays a critical role in the development of tolerance, the prevention of autoimmunity, and in anti-inflammatory responses. TGF-β2 is a potent inhibitor of intestinal epithelial cell (IEC) growth and stimulates IEC differentiation. The objective of this study was to determine whether a diet containing TGF-β2 modulates intestinal injury and immune responses in an Interleukin-10 knockout (IL-10−/−) mouse model of IBD. Methods: Five-week-old IL-10−/− mice (in BALB/c background) reared in our transgenic facility were fed either an enteral diet (Diet-A) containing TGF-β2 or a control enteral diet (Diet-B) not rich in TGF-β2. Mice were weighed weekly, monitored for illness and euthanized after eight weeks on the diet. Results: Final weights were 28 ± 1.2 g (58.2% gain) for Diet-A mice and 23 ± 1.6 g (32.9% gain) for Diet-B mice (p = 0.0194). The hematocrits were 48.3% for Diet-A compared to 42% for Diet-B mice (p = 0.0021). Mice on Diet-A had significantly lower serum TNF-α concentrations. Forty-four percent of mice on Diet-B developed severe diarrhea and rectal prolapse compared with none on Diet-A. Evaluation of intestinal pathology (score 0–4) revealed that animals fed Diet-A had a score of 2.1 ± 0.4 compared to 3.2 ± 0.36 in the Diet-B group (p = 0.040). The acute phase protein, serum amyloid A (SAA), was 3.8 times higher in the Diet-B group (p = 0.0038). Conclusions: IL-10−/− mice fed a TGF-β2 containing diet gained more weight, did not develop diarrhea or prolapse, had lower pathological scores, and lower SAAs. These data further support the use of TGF-β2 containing enteral diets as one mode of therapy for Crohn’s disease.

Immunohistochemical demonstration of alpha‐1‐proteinase inhibitor in brain tumors

Using light microscopy and immunoperoxidase methods (PAP), the presence of alpha‐1‐proteinase inhibitor (API) was studied in seventeen brain tumors and four normal brain samples. The brain tumors included four glioblastomas, five low‐grade gliomas, two metastatic lung carcinomas, two acoustic schwannomas, and four meningiomas. Normal brain displayed a finely granular intracytoplasmic staining confined to neuronal cells. Glial cells were negative for API. Fifteen of the 17 brain tumors were positive for API. Only two of five low‐grade gliomas were negative for API. Glioblastoma and metastatic tumors exhibited the strongest positivity followed by acoustic neuroma, meningioma, and low‐grade glioma. All positive samples exhibited finely granular intracytoplasmic API, and 50% exhibited extracellular API positivity. Metastatic and glioblastoma tumors demonstrated prominent extracellular API staining. Our results support the concept of a local production of API by brain tumors.

Herpes Simplex Virus: Fetal and Decidual Infection

We report 2 cases of probable in-utero transmission of herpes simplex virus. Herpes viral antigen was localized in these cases to the decidualized endometrium, using an immunoperoxidase staining technique. Routine histology did not show herpes inclusions in the decidual or glandular cells. This finding suggests that herpes virus may infect endometrial cells during pregnancy.

Identification of alpha-1-proteinase inhibitor-containing cells in pancreatic islets

SummaryBy a regular immunoperoxidase method, alpha-1-proteinase inhibitor (Api) was demonstrated in pancreatic islets in individuals with and without genetic deficiency of Api. Subsequently a double immunoperoxidase method, with two different chromogens (diaminobenzidine-brown and 4-chloro-1-naphthol-blue), was applied on the same tissue section in order to identify cells containing Api and cells secreting polypeptide hormones. Api-positive cells and hormone-secreting cells were found to be mutually exclusive indicating that Api is synthesized by previously unrecognized islet cell. The population of Api cells was significantly higher in persons with genetic deficiency than in other individuals, implying a possible compensatory hyperplasia of those cells triggered by a low level of circulating Api.