Adult bone marrow stromal cells differentiate into neural cells in vitro.

Bone marrow stromal cells (BMSC) normally give rise to bone, cartilage, and mesenchymal cells. Recently, bone marrow cells have been shown to have the capacity to differentiate into myocytes, hepatocytes, and glial cells. We now demonstrate that human and mouse BMSC can be induced to differentiate into neural cells under experimental cell culture conditions. BMSC cultured in the presence of EGF or BDNF expressed the protein and mRNA for nestin, a marker of neural precursors. These cultures also expressed glial fibrillary acidic protein (GFAP) and neuron-specific nuclear protein (NeuN). When labeled human or mouse BMSC were cultured with rat fetal mesencephalic or striatal cells, a small proportion of BMSC-derived cells differentiated into neuron-like cells expressing NeuN and glial cells expressing GFAP.

The X-gal caution in neural transplantation studies.

Cell transplantation into host brain requires a reliable cell marker to trace lineage and location of grafted cells in host tissue. The lacZ gene encodes the bacterial (E. coli) enzyme beta-galactosidase (beta-gal) and is commonly visualized as a blue intracellular precipitate following its incubation with a substrate, "X gal," in an oxidation reaction. LacZ is the "reporter gene" most commonly employed to follow gene expression in neural tissue or to track the fate of transplanted exogenous cells. If the reaction is not performed carefully-with adequate optimization and individualization of various parameters (e.g.. pH, concentration of reagents, addition of chelators, composition of fixatives) and the establishment of various controls--then misleading nonspecific background X-gal positivity can result, leading to the misidentification of cells. Some of this background results from endogenous nonbacterial beta-gal activity in discrete populations of neurons in the mammalian brain; some results from an excessive oxidation reaction. Surprisingly, few articles have empha sized how to recognize and to eliminate these potential confounding artifacts in order to maximize the utility and credibility of this histochemical technique as a cell marker. We briefly review the phenomenon in general, discuss a specific case that illustrates how an insufficiently scrutinized X-gal positivity can be a pitfall in cell transplantation studies, and then provide recommendations for optimizing the specificity and reliability of this histochemical reaction for discerning E. coli beta-gal activity.

Oxidative DNA damage in the aging mouse brain.

The brain exhibits regional vulnerabilities to many insults, and age itself has differential effects on neuronal populations as exemplified by the age-dependent loss of dopaminergic neurons in the nigrostriatal system. We hypothesized that oxidative damage to DNA was more likely to occur in the nigrostriatal system which undergoes significant neurochemical and functional changes with age. To test this hypothesis, oxidative damage to DNA, indicated by levels of 8-hydroxy2'-deoxyguanosine (oxo8dG), was measured in pons-medulla (PM), midbrain (MB), caudate-putamen (CP), hippocampus (HP), cerebellum (CB), and cerebral cortex (CX) at 3, 18, and 34 months of age in C57/b1 mice. Steady-state levels of oxo8dG increased significantly with age in MB, CP, and CB, but not in PM, HP, or CX. Manganese superoxide dismutase (MnSOD) activity decreased with age in MB, CP, and HP, but not in PM, CB, or CX. Regional activities of Cu/Zn superoxide dismutase (Cu/Zn SOD) and glutathione peroxidase (Glut Px) did not change significantly with age. Concomitant with the regional alterations in DNA damage, there was a significant age-dependent decline in locomotor activity, motor coordination, and striatal dopamine content especially during the interval between 18 and 34 months. In conclusion, oxyradical-associated damage to DNA did not accumulate uniformly across brain regions with age and was highest in brain regions that subserve spontaneous locomotor activity and motor coordination.

Fundoplication and gastrostomy in familial dysautonomia.

Fundoplication with gastrostomy has become a frequent treatment for patients with familial dysautonomia, so we evaluated the use of both procedures in 65 patients. Although patients differed widely in presenting signs and age, from 5 weeks to 40 years, gastroesophageal reflux was documented in 95% of patients by cineradiography or pH monitoring. Panendoscopy was a useful adjunct. Preoperative symptoms of gastroesophageal reflux included vomiting, respiratory infections, and exaggerated autonomic dysfunction. Severe oropharyngeal incoordination frequently coexisted and resulted in misdirected swallows with aspiration, dependence on gavage feedings, or poor weight gain and dehydration. Follow-up after surgical correction ranged from 3 months to 11 years; 55 patients (85%) were available for a 1-year postoperative assessment. We had no instances of surgical death. The long-term mortality rate was 14%, primarily related to severe preexisting respiratory disease. Beyond the first postoperative year, 30 patients had pneumonia attributed to continued aspiration, exacerbation of preexisting lung disease, or recurrence of gastroesophageal reflux. Of 11 patients who vomited postoperatively, six had recurrence of reflux. Recurrence of gastroesophageal reflux was documented in eight patients (12%), and we revised the fundoplication in three patients. The number of patients with cyclic crises was reduced from 18 to 7; retching replaced overt vomiting in all but two of these seven patients, neither of whom had recurrence of reflux. Because oropharyngeal incoordination was prominent, concomitant use of gastrostomy and an antireflux procedure was especially effective in the treatment of younger patients with familial dysautonomia, before the development of severe respiratory disease. Despite the development of severe morning nausea in 15 patients, the combination procedure resulted in significantly improved nutritional status, decreased vomiting, and decreased respiratory problems. Appropriate use of gastrostomy feedings also contributed to success of the operation. The generally good outcome of fundoplication with gastrostomy confirms the benefit of this procedure in familial dysautonomia.

Perirectal abscess in the Hermansky-Pudlak syndrome.

The Hermansky-Pudlak syndrome (HPS) is a triad of tyrosine-positive albinism, platelet dysfunction, and the deposition of an abnormal ceroid-like pigment in the tissues. Complications of the syndrome, such as pulmonary fibrosis, renal failure, and cardiomyopathy, have been described. Granulomatous colitis has been documented in several families with the HPS. The bowel disease of the HPS is a unique type of inflammatory bowel disease with clinical features suggestive of idiopathic ulcerative colitis and pathologic features suggestive of Crohn's disease. Analogous to the presentation of Crohn's disease with perianal and perirectal involvement, we describe the occurrence of perianal disease and a perirectal abscess in a 29-yr-old woman with HPS and mild granulomatous colitis.

Bradycardia associated with hiatal hernia and gastroesophageal reflux relieved by surgery.

A man known to have familial dysautonomia presented with a cardiac arrhythmia due to development of hiatal hernia and gastroesophageal reflux. Preoperative symptoms and assessment are described including use of power spectrum analysis of heart rate fluctuations which was consistent with enhanced parasympathetic stimulation. After surgical repair of hiatal hernia and fundoplication, bradycardia resolved, gastroesophageal reflux symptoms subsided, and the power spectrum analysis of heart rate confirmed decreased parasympathetic influence. Power spectrum analysis proved to be a useful adjunct in confirming preoperative autonomic imbalance and assessing the postoperative result. It is concluded that in individuals with disorders such as familial dysautonomia that are associated with autonomic dysfunction, cardiac arrhythmias may be a sign of esophageal pathology. Thus, cardiac evaluations should be accompanied by investigation of gastroesophageal structure and function and appropriate treatment may prevent a catastrophic arrhythmia.

Carcinoma of the small intestine in regional enteritis: presentation of a case and review of the literature.

A case of carcinoma of the jejunum occurring in a patient with Crohn's disease of 30-years' duration is presented. Forty-seven previously reported small bowel carcinomas in Crohn's disease patients are reviewed. The incidence of small bowel carcinomas in patients with Crohn's disease has been cited as being as low as 0.08%, a figure which is still higher than the 1 in 10(9) population which would be predicted if the two diseases were independent of one another. The average age of incidence of the Crohn's carcinomas was 46.5 years while that for the de novo group was 55 years. The sexual ratios were 2.46:1 and 2:1, males to females, for the respective groups. The de novo carcinomas had a slight predilection for the duodenum at 40.7% while the latter group had a heavy predilection for the ileum at 70.8% and contained no duodenal carcinomas. The prognosis of the Crohn's group appeared to be much worse than that of the de novo group with five-year survivals of 3.7% and 20-22% respectively. Late diagnosis in the enteritis group was felt to be the major reason for this. Finally, several pathological differences between the lesions in both groups are presented.

A clinical and laboratory evaluation of immune serum globulin from donors with a history of hepatitis: attempted prevention of post-transfusion hepatitis.

A controlled trial of passive immunization for prevention of post-transfusion viral hepatitis was carried out in order to determine whether effective levels of antibody were present in the "convalescent" immune serum globulin used in the study. This globulin was prepared selectively from plasma of donors giving a history of overt viral hepatitis two or more years earlier. The proportion of contributors to the globulin who had B hepatitis was unknown but the final product contained a low titer of antibody to the surface antigen of hepatitis B virus (anti-HBs). The failure of 20 ml of immune serum globulin to reduce the incidence of type B post-transfusion hepatitis (7/93) below that of placebo-treated controls (8/102) was not unexpected in view of the globulin's low titer of anti-HBs. However, more than two thirds of the post-transfusion cases were not type B and were as plentiful among globulin recipients (17/93) as among controls (17/102). Although some of the donors from whom the immune serum globulin was obtained may once have had the same type(s) of hepatitis as the non-B cases currently observed in transfusion recipients, the globulin apparently did not contain enough specific antibody to confer protection in the dose schedule tested.

The radiology corner: the small bowel in immunoglobulin deficiency syndromes.

Recent advances in immunology have permitted recognition of a group of patients who have gastrointestinal manifestations as part of an immunoglobulin deficency syndrome. Such immunoglobulin deficiency may be primary or may be secondary to a variety of diseases. We have classified and described the small bowel roentgen features associated with the various immunoglobulin deficiency syndromes as follows: 1. the sprue pattern, as seen in hypogammaglobulinemic sprue and in Ig-A deficient sprue; 2. multiple nodular defects; 3. inflammatory changes secondary to giardiasis, associated with immune deficiency diseases; 4. thickening of the small intestinal folds, as seen in the plasma cell dyscrasias, lymphoma, intestinal lymphangiectasia and amyloidosis.