DR.lyp/lyp bone marrow maintains lymphopenia and promotes diabetes in lyp/lyp but not in +/+ recipient DR.lyp BB rats

Lymphopenia is due to a frameshift mutation in Gimap5 on rat chromosome 4 and is linked to type 1 diabetes in the diabetes prone (DP) BB rat. The hypothesis that bone marrow derived cells confer the lymphopenia phenotype was tested by reciprocal bone marrow transplantation in 40-day-old lethally irradiated diabetes resistant (DR) congenic DR.lyp/lyp (lymphopenia and diabetes) and DR.+/+ (no lymphopenia and no diabetes) rats. In two independent series of transplants, all DR.lyp/lyp rats (n=5 and 4) receiving DR.lyp/lyp bone marrow retained lymphopenia and developed insulitis (5/5 and 4/4) as well as diabetes in some (2/5 and 3/4). Both DR.+/+ and DR.lyp/lyp rats receiving DR.+/+ bone marrow cells as well as DR.+/+ rats receiving DR.lyp/lyp bone marrow cells showed no lymphopenia or diabetes. In accordance with earlier studies in non-congenic BB rats, the DR.+/+ rats receiving DR.lyp/lyp bone marrow cells recapitulated an intermediary phenotype rather than the +/+ or lyp/lyp phenotypes. Our data demonstrate that BBDP rat lymphopenia and diabetes are transferred by bone marrow transplantation to syngeneic DR.lyp/lyp but not DR.+/+ recipients. The intermediary recapitulation of DR.lyp/lyp T cells in recipient DR.+/-/+/- rats suggests that radiation resistant +/-/+/- T cells, the Gimap5 mutation in bone marrow cells, or both may not support the development of lymphopenia.     

FK-506 prevents diabetes in diabetes-prone BB/Wor rats.

The effect of the immunosuppressant FK-506 on the development of diabetes in BB/Wor rats was investigated. Using a treatment schedule (25 micrograms i.m. from day 27 to 120), not associated with detectable general ill effects, this drug was found to completely inhibit the appearance hyperglycemia and to reduce the histological signs of pancreatic insulitis. The treatment was also able to reduce the percentages of Ia+ T-lymphocytes and to block the appearance of detectable serum levels of gamma interferon (IFN).     

煅烧骨/壳聚糖复合材料促进SD大鼠骨髓间充质干细胞的成骨分化

背景:课题组前期研究已经证实煅烧骨/壳聚糖复合材料的微观结构与自然骨组织相似,安全无毒,抗压强度良好,并且能够促进成骨细胞的增殖及黏附,因此继续研究其对骨髓间充质干细胞成骨分化的作用具有重要意义。目的:探讨煅烧骨/壳聚糖复合材料对骨髓间充质干细胞成骨分化的影响。方法:将煅烧骨/壳聚糖复合材料与SD大鼠骨髓间充质干细胞体外共培养并进行成骨诱导,成骨诱导1,2,3周时,采用实时荧光定量PCR检测成骨分化相关基因的表达。结果与结论:煅烧骨/壳聚糖复合材料能促进骨髓间充质干细胞的增殖黏附,并促进成骨相关基因碱性磷酸酶、Ⅰ型胶原、骨钙素的表达,具有诱导骨髓间充质干细胞成骨分化的作用。     

Varicella zoster gastritis in a bone marrow transplant recipient.

A case is reported of a patient who had previously undergone autologous bone marrow transplantation for recurrent Hodgkin's disease. The patient developed a generalised vesicular skin eruption. The clinical diagnosis was of disseminated shingles. Herpes viral particles were identified within the vesicular fluid by electron microscopy and using a specific monoclonal antibody to varicella zoster virus (VZV), positive immunofluorescence was detected in scrapings from the base of a vesicle. Gastroscopy and biopsy were performed because of severe abdominal pain and vomiting. The histological features were of non-specific active inflammation. Despite the histological absence of viral inclusions electron microscopy of the gastric biopsy revealed the presence of intranuclear herpes viral particles with a diameter of 90-100 nm. VZV specific DNA was detected by the polymerase chain reaction in the gastric biopsy extract. The patient was treated with acyclovir and made a full recovery.     

Intermittent feeding and fasting reduces diabetes incidence in BB rats.

Food intake may be one of several factors which influence the risk of development of insulin dependent diabetes mellitus, but the influence of the pattern of food supply has not been studied previously. The aim of the present study was to investigate the effect of intermittent feeding and fasting upon diabetes in BB rats. This study included three groups. Group 1 served as control and included 77 animals, 79% became diabetic. In groups 2 and 3, after weaning, food but not water was withdrawn from the animals: 24 h twice a week in group 2; 24 h every second day in group 3. Group 2 included 40 BB rats, 50% (p < 0.004) became diabetic. Group 3 included 44 BB rats, 52% (p < 0.01) became diabetic. No differences were seen between sexes. Degree of insulitis was not influenced by changed food supply. Regarding blood glucose, no influence was seen among diabetic animals, among non-diabetic animals changed food supply reduced blood glucose values obtained at the end of the study. Intermittent feeding and fasting tended to reduce mean age at the time of diagnosis of diabetes, significance was reached only in female animals from group 3 compared to group 1. Body weight was obtained weekly. Intermittent feeding and fasting caused a reduced weight gain in group 2 as well as in group 3 compared to control animals; however, most pronounced in group 3 and also more pronounced among males compared to females. For pre-diabetic and non-diabetic animals comparable influence on body weight was seen. The main conclusion in the study is that intermittent feeding and fasting reduced diabetes incidence in BB rats.     

Fatal adenovirus 32 infection in a bone marrow transplant recipient.

A case of disseminated adenovirus type 32 infection causing severe hepatitis, gastrointestinal ulceration and also with respiratory involvement is reported in a bone marrow transplant recipient. Typical viral inclusions were seen in the postmortem histological sections and adenovirus infection was confirmed using in situ hybridisation and isolation of adenovirus type 32 from separate organs at necropsy. This is the first case in which adenovirus 32 was the cause of fatal disseminated disease in a bone marrow transplant recipient.     

Paecilomyces lilacinus fungemia in an adult bone marrow transplant recipient.

Paecilomyces lilacinus is a rare fungal pathogen in humans. We report a case of fungemia caused by P. lilacinus in a non-neutropenic adult, 120 days after bone marrow transplant. The patient's primary risk factor was the presence of an indwelling vascular catheter. Her initial clinical course was characterized by fever, chills, and rigors. Blood cultures from the central line and peripheral veins were positive, as was a peripheral specimen drawn after removal of the catheter. Two initial peripheral specimens were positive for P. lilacinus only by blind subculture and/or sustained incubation. She developed peripheral pulmonary nodules following the fungemia, thus raising the possibility of disseminated disease, but definitive diagnosis was confounded by Pseudomonas bacteremia. The nodules cleared and she recovered following removal of the central line and treatment with amphotericin B and 5-fluorocytosine, despite in vitro resistance to these antifungal drugs. This case underscores the increasing importance of P. lilacinus as a human pathogen capable of producing disease in immunocompetent, as well as in immunocompromised hosts. Also of note is that blood culture systems may require extended incubation or subcultures in order to detect fungi.     

Brain-reactive autoantibodies in BB/d rats do not recognize glutamic acid decarboxylase.

The BB rat spontaneously develops insulin-dependent diabetes mellitus (IDDM) similar to that in humans. The most practical markers of beta cell autoimmunity are circulating antibodies to islet cell components. In particular autoantibodies to the enzyme glutamic acid decarboxylase (GAD) are a common feature of IDDM development in humans. This study aims at investigating the prevalence and levels of autoantibodies in BB rats to antigens in a semipurified, GAD-enriched preparation from rat brain. Eighteen diabetes-prone BB/d rats (10 male and eight female) were tail bled weekly from age 28 days to 113 days and antibodies detected on the rat brain preparation by ELISA. Antibody levels were expressed as arbitrary units relative to a standard positive serum. Individual rats varied in the time and order of antibody appearance and IDDM onset, with the earliest occurrence being 42 days and 69 days, respectively. In some rats antibody production was maintained but declined in others. By 113 days 85% of diabetic rats had at some time been positive for autoantibodies to brain components, compared with 25% of non-diabetics (P = 0.09 by Fisher's exact test). Immunoabsorption studies using recombinant rat GAD-65 or recombinant human GAD-67 failed to inhibit the binding of BB rat sera to the original rat brain preparation. A capture ELISA using GAD-6 MoAb to capture GAD-65 from rat brain preparation or from a preparation of recombinant rat GAD-65, failed to detect anti-GAD antibodies in BB rats. Immunofluorescent staining of tissue sections showed the autoantibodies to be brain-specific, but having distinct staining patterns to the anti-GAD antibodies of Stiff Man Syndrome serum. In conclusion, BB rats possess autoantibodies reactive with rat brain antigens which may be associated with IDDM. However, these are not directed against GAD.     

Disseminated Microascus cirrosus infection in pediatric bone marrow transplant recipient.

Microascus cirrosus Curzi and its associated anamorphic state, Scopulariopsis, were recovered from the cutaneous lesion of a 12-year-old male who had undergone an autologous bone marrow transplantation for acute myelogenous leukemia. Histopathology sections from the biopsied lesion demonstrated septate hyphae consistent with a fungal etiology. Radiographic studies of the lungs subsequent to progression of the lesion revealed a consolidation in the right upper lobe suggesting a primary focus of infection. While M. cirrosus is fairly abundant in nature and widely distributed in stored grains in North America, this is, to our knowledge, the first reported human infection by this species. Salient characteristics for the identification of this dematiaceous ascomycete, M. cirrosus, will be presented.     

The gene for the T lymphocyte alloantigen, RT6, is not linked to either diabetes or lymphopenia and is not defective in the BB rat

In an outcross between a diabetic BB/H rat and a healthy Long Evans Hooded rat, the segregation of the RT6 gene was studied in the 207 F2 animals to look for linkage with diabetes or lymphopenia. The recessive gene, albino (c), was used as a marker for the RT6 gene because of the close proximity of these two genes on chromosome 1. Though most of the albino F2 rats should have been homozygous for the BB RT6 gene, we found no increase in the incidence of diabetes or lymphopenia among them when compared to their hooded littermates. Therefore, the RT6 gene was not linked to diabetes or lymphopenia in the BB rat. Moreover, the non-lymphopenic albino rats displayed normal RT6 expression when compared to the normal hooded rats showing that the RT6 gene from the BB/H grandfather was not defective. Any alteration in lymphocyte composition which could be specifically related to diabetes was studied by measuring all F2 rats for the major lymphocyte subsets including the RT6+ subset. We found that the typical pattern of lymphopenia described in diabetic BB rats was displayed by both diabetic and non-diabetic lymphopenic rats in the F2 generation. Thus, all these lymphocyte abnormalities including the depletion in RT6+ T lymphocytes appeared as a consequence of lymphopenia alone and could not be specifically related to diabetes.     

Anti-diabetogenic effect of fusidic acid in diabetes prone BB rats.

Fusidic acid and its sodium salt (fusidin) are anti-staphylococcal drugs. In vitro studies have shown that they prevent the lymphocyte co-stimulatory activities of the cytokines IL-1 and IL-6 in a manner similar to that of cyclosporin A, and prevent the inhibitory effect of IL-1 on glucose-induced insulin production. As IL-1 and IL-6 are thought to play a role in the pathogenesis of Type 1 diabetes, the aim of this study was to investigate whether fusidin could influence the disease incidence of the spontaneously diabetic BB rat model. Accordingly, a group of 50 BB rats receiving fusidin dissolved in their drinking water were compared to a control group of 55 rats over a period of 200 days. The incidence of diabetes was found to be 52% in the experimental group and 71% in the control group (P < 0.05). The degree of insulitis and the number of islets at histological examination were similar among the non-diabetic animals whereas the diabetic fusidin-treated animals showed a higher degree of islet preservation than the diabetic control rats. The results are highly indicative of an anti-diabetogenic effect of fusidin.     

Long-term regular exercise promotes memory and learning in young but not in older rats

BACKGROUND AND OBJECTIVE: By aging, some functions in nervous system like spatial memory are reduced. It has been shown that short-time physical activity can improve memory but there is much less data on the long-term exercising. In the present study, the aim was to clarify the effect of regular long-term physical activity on spatial memory and learning of young and middle aged and older male Wistar rats. MATERIALS AND METHODS: Sixty 3 months old rats were randomly divided in six equal groups. Experimental groups were treadmill exercised at speed 22m/min for 1h 6 days per week, and the program lasted 3, 6 and 9 months, respectively. At the end of training period, spatial memory of rats was tested using Morris Water Maze. RESULTS: Results indicated that regular physical activity significantly increased spatial memory (p<0.05) in young rats (6 months old) as compared to controls, but not in the older ones (9 and 12 months old). Nonetheless, spatial memory of these rats was significantly better than in younger ones in both groups (p<0.05). CONCLUSION: Spatial memory and learning increased due regular exercise in young rats. With progression of age up to 9 and 12 months the memory improved, but the programmed exercise had no positive effect on learning.     

Catheter-related Rahnella aquatilis bacteremia in a pediatric bone marrow transplant recipient.

Rahnella aquatilis, a rarely encountered member of the family Enterobacteriaceae, was twice isolated from the blood of a pediatric bone marrow transplant recipient. This is the first report of a pediatric case of R. aquatilis bacteremia, and it was probably related to inappropriate handling of a Hickman catheter.     

Replacement of recipient stromal/mesenchymal cells after bone marrow transplantation using bone fragments and cultured osteoblast-like cells.

Abstract We present our experience on treatment of three children with potentially fatal diseases using a unique protocol for non-myeloablative bone marrow transplantation. The protocol was designed to promote engraftment of bone marrow stromal/mesenchymal cells (SC/MSCs) based on the knowledge from preclinical models over the last three decades. Accordingly, our protocol is the first to test the use of bone fragments as an ideal vehicle to transplant such cells residing in the bone core. Because of the paucity of knowledge for optimum transplantation of SC/MSCs in humans, we used a multifaceted approach and implanted bone fragments both intraperitoneally and directly into bone on day 0 of BMT. We also infused cultured donor osteoblast-like cells intravenously post-BMT. We were able to achieve high levels of stroma cell engraftment as defined by molecular analyses of bone biopsy specimens.     

Liver perisinusoidal fibrosis in BB rats with or without overt diabetes.

Perisinusoidal fibrosis is a vascular lesion observed in the liver of type I diabetic patients. To investigate whether this liver lesion is secondary to hyperglycemia or whether it represents a separate collagen vascular disorder, the authors studied the structure of liver sinusoids in genetically susceptible BB rats in which a spontaneous diabetes develops similar to human type I diabetes. Seven diabetic insulin-treated BB rats, 7 nondiabetic BB rats, and 6 control non-BB rats were studied. Histologic abnormalities of the collagen network were detected on trichrome-stained sections. Perisinusoidal collagen fibers were quantified ultrastructurally by the point-counting method. All control non-BB rats had normal livers; 86% of the diabetic as well as 71% of the nondiabetic BB rats displayed localized sinusoidal thickening corresponding ultrastructurally to perisinusoidal fibrosis; in these abnormal rats the percentage of collagen fibers per sinusoid unit was significantly higher than that in controls. Fibrous septa (2 diabetic and 5 nondiabetic BB rats) and liver nodulation (3 diabetic and 1 nondiabetic BB rats) were also observed. Perisinusoidal fibrosis is a frequent liver vascular abnormality in a strain of rats genetically predisposed to the development of type I diabetes. The lesion is independent of the presence of diabetes. These observations suggest that liver perisinusoidal fibrosis in patients with type I diabetes might be linked to a genetic abnormality rather than to hyperglycemia per se.     

Disseminated, fatal Trichosporon asahii infection in a bone marrow transplant recipient.

Trichosporon asahii is the most important species regularly isolated from systemic mycoses and shows a predilection for hematogenous dissemination. This report describes the first fatal case of disseminated trichosporonosis caused by T. asahii in a patient with familial aplastic anemia (AA). An 11-year-old girl with familial AA received chemoradiotherapy and immunosuppressive therapy for bone marrow transplantation. She was neutropenic and suffered from fever, cough, and severe mouth ulcers. T. asahii was repeatedly demonstrated by appropriate morphological and physiological characteristics, i.e., arthroconidium formation, urease activity, and assimilation of carbon and nitrogen compounds. T. asahii was found in samples of sputum, nose, and mouth ulcers by direct microscopy and culturing. Furthermore, postmortem histopathology study revealed vast tissue invasion of fungal hyphae characteristic of Trichosporon in the lung and liver. Disseminated trichosporonosis should be suspected in immunocompromised patients when a febrile condition does not improve after prolonged treatment with broad-spectrum parenteral antibiotics.     

Effect of gliotoxin on development of diabetes mellitus in diabetes-prone BB/Wor rats

The object of this investigation was to determine if gliotoxin, an immunomodulating fungal secondary metabolite, is capable of preventing the development of autoimmune diabetes mellitus in diabetes-prone BB/Wor rats. Chronic treatment, consisting of 1 microg gliotoxin/g of body wt administered three times weekly from the age of 30 days through 120 days, reduced the incidence of diabetes from 90% diabetic by 120 days among vehicle-treated animals to 56% diabetic among gliotoxin-treated animals. This result was significant by life table analysis. Animals treated with gliotoxin maintained lower serum glucose levels even in the pre-diabetic state than control (vehicle-treated) rats. Gliotoxin at levels used in this study showed no appreciable effect on the viability of rat insulinoma (RIN 38) cells in culture and only slightly decreased their insulin secretion. Animals chronically treated with gliotoxin showed weight gains comparable to those seen in controls, and the effect of gliotoxin on peripheral blood leukocyte counts was not significant. The possibility that gliotoxin exerted its effect through immunomodulating effects was implied by the loss of white pulp in splenic follicles of gliotoxin-treated animals.     

Necrotizing cerebritis in an allogeneic bone marrow transplant recipient due to Cladophialophora bantiana.

We describe a necrotizing cerebritis in an allogeneic bone marrow transplant recipient caused by the neurotropic, dematiaceous fungus Cladophialophora bantiana. The patient presented 7 months after bone marrow transplantation with fever and sudden onset of left-sided weakness, followed shortly by cranial nerve III and VI palsies. The patient had a lesion (3.0 by 2.0 by 2.0 cm) of the right midbrain with extension to the pons, the left brain stem, and the right superior and the middle cerebellar peduncles. The diagnosis was made by microscopic examination and culture of a brain biopsy.