Systemic cell-mediated reactions in vivo. Effect of the interaction of circulating antigen with sensitized lymphocytes on glomeruli and pulmonary alveoli

The effects of systemic cell-mediated hypersensitivity reactions on glomeruli and lungs were investigated in rats. The animals were given an intravenous injection of antigen 7 days after sensitization or were given an intravenous injection of lymph node cells from sensitized syngeneic donors 1 day after antigen injection. Control animals were given an irrelevant antigen or saline. All animals received three injections of 3H-thymidine during the course of the experiments. The animals were sacrificed 2 or 3 days after antigen injection. Autoradiographs of renal and pulmonary tissue showed significantly more labeled mononuclear cells in glomeruli and pulmonary alveolar walls in the experimental groups than in the control groups. Immunofluorescence studies did not reveal antigen, rat IgG, or C3 in glomeruli. The results indicate that systemic cell-mediated reactions can lead to an accumulation of mononuclear cells in glomeruli and lungs, an effect that may contribute to tissue injury.     

del(16)(q22) in a child with acute myeloid leukemia without bone marrow eosinophilia.

In a study comprised of 79 cancer patients selected for determining chromosomal damage to peripheral blood lymphocytes before, during, and after radiotherapy, four patients revealed chromosome aberrations prior to radiotherapy in 5% of the cells examined. The most common anomalies were dicentric chromosomes and translocations of unidentified chromosomal material on the long or short arm. We attribute this high incidence of chromosomal aberrations before radiotherapy to a combination of various external factors that are known to cause such cytogenetic injury.     

Coincidence of cytogenetic markers in four murine cell lines

Foreign body tumorigenesis was induced by the subcutaneous implantation of a plastic or glass cylinder in BALB/c mice; the inoculation of human neoplastic cells significantly increased the incidence of these anaplastic sarcomas. Of 15 tumors studied, four presented the same markers: one induced with and three without human neoplastic cell inoculation within the foreign body. The markers observed were double minutes (DM), a long acrocentric marker (MLA), and a metacentric marker (MM). The DM are a number of small often tiny chromosomal structures appearing in pairs together with chromosomes of ordinary size. MLA is a long acrocentric derived from a translocation in tandem between chromosomes #1 and #16. MM is due to centric fusion of two chromosomes #10. Numerical anomalies consisted of gains of the same chromosomes types. It is postulated that these coincident findings are related to the foreignbody tumorigenesis.     

Cytogenetic changes in hepatocarcinomas from rats treated with chronic exposure to diethylnitrosamine.

Cytogenetic analysis of rat hepatocarcinomas obtained after diethylnitrosamine (DEN) exposure showed a wide variety of numerical and structural chromosomal changes: 53 of 86 hepatocellular carcinomas showed at least one recurrent chromosomal aberration. Some of these recurrent changes occurred in several tumors. Chromosomes 1, 3, 11, and 12 were abnormal in more than 30% of the carcinomas; chromosomes 2, 4, 5, and 10 were abnormal in 10%. Moreover, chromosomes 1 and 10 were generally lost or deleted and chromosome 3, 4, and 11 were very often gained. The most frequent anomaly was loss of chromosome 1 which was observed in 35% of the tetraploid cell populations. The occurrence in several tumors of recurrent chromosomal rearrangements as well as various repeated aneuploidies strongly suggests that these anomalies are implicated in the process of rat hepatocarcinogenesis induced by DEN treatment.     

Toxopathological and cytogenetic effects of aflatoxin B1 (AFB1) on pregnant rats

The present study was carried out to evaluate the toxopathological effects and macro-DNA damage of Aflatoxin B1 (AFB1) in pregnant rats at a dose of 1 mg/kg. body wt., given from 6th to 15th day of gestation. The effects and damage are represented by histopathological changes and different types of chromosomal aberrations in dams, in addition to teratogenic changes in the feti. Pregnant dams revealed a significant decrease in their body weights and gross enlargement of the liver. Histologically, the liver showed necrotic areas and congested central vein. The kidneys revealed interstitial hemorrhages, renal casts, degeneration and necrosis. The lungs revealed lymphocytic infiltrates in the interstitial tissue, while the spleen revealed lymphoid depletion. Chromosomal analysis revealed both structural and numerical chromosomal aberration, including centromeric attenuations, chromatid gaps, chromatid breaks, end-to-end associations, fragments, ring chromosomes, deletions, dicentric chromosomes, chromosomal fusions, centric fusions, stickness and hypoployploidy. Centromeric attenuations and end-to-end associations were more frequent than other chromosomal aberrations. Concerning the teratogenic effects in the fetuses, the toxin induced multiple skeletal anomalies. These anomalies included incomplete ossification of skull bones and failure of ossification of long and flat bones.     

Random X-inactivation in a girl with duplication Xp11.21–p21.3: Report of a patient and review of the literature

We describe a 10-month-old girl with abnormal clinical findings and Xp duplication. She showed poor weight gain and developmental retardation, and had several minor anomalies including pigmentary dysplasia (hypomelanosis of Ito). She had a partial short arm duplication in the paternally derived X chromosome, 46,X,dup(X)(p11.21p21.3), with the normal and duplicated X chromosomes randomly inactivated. These findings indicate that gross functional imbalance in the cells with an active dup(X) chromosome has caused global developmental defects in the patient, and that functional chromosomal mosaicism with respect to the duplicated Xp region has resulted in pigmentary dysplasia. Literature review of 52 patients with partial X duplications revealed (1) random or skewed but not completely selective X-inactivation in 9 of 45 patients examined for the X-inactivation pattern, independently of the size or location of duplicated segments, (2) apparently normal phenotype in 6 of 9 patients with random or skewed X-inactivation, and (3) an abnormal phenotype in 13 of 35 patients with completely selective inactivation of dup(X) chromosomes. Am. J. Med. Genet. 86:44–50, 1999. © 1999 Wiley-Liss, Inc.     

Chromosomal aberrations in infective hepatitis

Structural chromosomal aberrations, in the form of breaks, were found in a significantly higher proportion of bone marrow cells in patients with infective hepatitis than in controls. These anomalies were observed during the first and third weeks after the onset of jaundice but had subsided by the sixth week.Chromosomal aberrations did not appear to be related to the severity of infective hepatitis or to the sex or age of the patients.The distribution of chromosomal abnormalities did not appear to be random; they were observed predominantly in the A(2) and B(4-5) series. Since no abnormalities were detected in the G-group chromosomes, no evidence in support of a relationship between infective hepatitis and Down's syndrome was obtained.Numerical chromosomal aberrations were not observed, nor was any evidence obtained that mitotic activity of bone marrow cells is suppressed in patients with infective hepatitis.     

Placental and fetal alterations due to Venezuelan equine encephalitis virus in rats.

Histopathological changes in the placentas, embryos, and fetuses of rats inoculated intraperitoneally with the virulent Guajira strain of Venezuelan equine encephalitis virus were studied by light microscopy and immunoperoxidase methods. Rats inoculated before day 15 of pregnancy showed necrosis and hemorrhages in the embryonic disks. Swelling of cytoplasm and nuclear pyknosis of cyto- and syncytotrophoblastic cells were noted as early as 2 days after inoculation. During weeks 1 and 2 of pregnancy, death of the embryos was always observed 3 to 4 days after Venezuelan equine encephalitis virus inoculation. Placental and fetal damage varied among the specimens. In rats 18 days pregnant and sacrificed 2 days after inoculation, there were some viable fetuses; the placentas showed inflammatory reactions in the mesometrial and decidual vessels. Other rats sacrificed at 3 to 4 days after inoculation showed large placental infarcts with fetal death. Viremia peaked during day 2 after inoculation. Immunoperoxidase stains demonstrated viral antigens present in the decidua, myometrium, and cyto- and syncytotrophoblastic cells. These experiments provide additional data regarding the pathogenesis and structural damage in the placental and fetal tissues caused by Venezuelan equine encephalitis virus.     

Bleomycin effects on mouse meiotic chromosomes.

The effects of a radiomimetic chemical, bleomycin (BLM), on meiotic chromosomes was evaluated in mice treated by intraperitoneal (i.p.) or intratesticular (i.t.) injection. Chromosome aberrations were analyzed at meiotic metaphase I, and damage to the synaptonemal complex (SC) was analyzed in meiotic prophase cells. In the metaphase aberration studies, an i.p. injection of 80 mg/kg BLM, timed to precede or coincide with pre-meiotic S phase, led to a significant increase in structural damage (P less than 0.01) in cells reaching metaphase I 12 days after treatment. However, no increases in clastogenic effects were observed at metaphase I after treatment of cells during various stages of prophase. SC analyses in pachytene cells following an i.p. or i.t. injection at S phase revealed various forms of synaptic errors and structural anomalies, including qualitative changes similar to those observed following irradiation. I.p. doses ranging from 25 to 100 mg/kg, and i.t. doses as low as 0.5 mg/kg, caused roughly 6-fold increases over control levels in the number of damaged cells. SC analyses in pachytene cells following BLM treatments 2 days earlier (at leptotene-zygotene) or 16 h earlier (at early-mid pachytene), also revealed induced structural and synaptic anomalies. Following the treatment at early-mid pachytene, there was some suggestion of interference with chiasma formation as evidenced by univalent-like configurations detected at diakinesis-metaphase. It was concluded that BLM is clastogenic for meiotic chromosomes; however, it does not reveal the strong S-independent clastogenic activity at meiosis that is characteristic of its activity at meiosis. SC analysis indicated that some damage is induced at meiotic prophase, although structurally aberrant cells are not recoverable at meiotic metaphase I. The results call forth various possible explanations for germ-line specific responses to BLM clastogenic activity.     

Synthesis and migration of 3H-fucose-labeled glycoproteins in the ciliary epithelium of the eye: Effects of microtubule-disrupting drugs

³H-fucose was injected intravenously or intravitreously into albino rats. After time intervals of 10, 40, and 50 min, 1, 1.5, and 4 hr, 1, 3, and 7 days, and 1, 2, and 4 weeks after injection, the animals were sacrificed by intracardiac perfusion with gluteraldehyde. Samples of the ciliary body were prepared for light and electron microscope radioautography. Light microscope autoradiographs showed that the cells of both the inner and outer layers of ciliary epithelium actively incorporated ³H-fucose label in a reaction that peaked in intensity at 4 hr after injection, and then progressively declined. Electron microscope radioautographs revealed that, at early time intervals, most of the label was localized to the Golgi apparatus. With time, the plasma membrane of both cell types became increasingly labeled, and accounted for 60–70% of the total silver grains at 4 hr after injection. Adjacent to the basal cell surface of the inner layer cells, the fibers of the zonula became increasingly labeled from 1.5 hr onwards, providing strong evidence that these cells secrete glycoproteins to the zonula. When vinblastine was administered 30 min before ³H-fucose injection, followed by sacrifice 1.5 hr later, a much larger proportion of label remained localized to the Golgi apparatus than in controls, and the plasma membrane and zonula were much less labeled. These results suggest that, as documented in other cell types, microtubules may play a role in the intracellular transport of membrane and secretory glycoproteins in these cells.     

The ultrastructure of the thyrotropic cell during thyrotropin rebound in the adenohypophysis of the rat

Adenohypophyses of rats were studied ultrastructurally to ascertain the morphologic changes that occur in thyrotropic cells during the pituitary TSH Rebound Phenomenon. Rats were maintained on propylthiouracil (PTU) for 43–147 days and their adenohypophyses studied three to nine days after discontinuance of goitrogen treatment. TSH rebound was also induced in chronically hypothyroid rats by single intravenous injections of 0.8, 4, 20 and 200 μg of thyroxine. Pituitaries were studied from animals sacrificed 6–24 hours after thyroxine injection. Thyrotrophs of euthyroid rats were characterized ultrastructurally by the presence of numerous peripherally-located, small secretory granules (storage phase) and by highly dilated cisternae of rough endoplasmic reticulum (secretory phase). The thyrotropic cells in PTU-treated rats were sparsely granulated, displayed enlarged mitochondria with much loss of cristae and contained extensively dilated rough endoplasmic reticulum and expanded Golgi membranes. Marked repletion of granules, both intracisternal and cytoplasmic, was seen during TSH rebound in the pituitary. It was observed in some thyrotrophs as early as three days after goitrogen withdrawal and was enhanced further at the 6- and 9-day intervals. Granule repletion increased progressively with doses of thyroxine up to 20 μg and assayable thyrotropin in pituitary glands increased 500–700% over normal. Effects with the 20 μg dose could be detected as early as six hours. With higher doses of thyroxine, granule repletion in thyrotrophs was inhibited and mitcohondrial hypertrophy was largely reversed. The electron microscopic findings reveal close correlation with previously published data on thyrotropin assays of the rat pituitary. They support the concept that the degree of cytoplasmic granulation in thyrotropic cells is quantitatively related to assayable stores of thyrotropin in the anterior pituitary gland.     

Mesothelial cell sheets cultured on fibrin gel prevent adhesion formation in an intestinal hernia model

In the present study, we examined a novel technique to prevent adhesion formation in a rat intestinal hernia model with mesothelial cell sheets cultured on fibrin gel. Mesothelial cells were obtained from isologous rats by enzymatic disaggregation of mesentery and cultured on fibrin gel. Electron microscopy revealed that these cultured cells form contiguous monolayer cell sheets with well-developed microvilli. These tissue-engineered constructs were grafted in vivo to an intestinal hernia model that results in regular surgical adhesions without treatment. Five days postgrafting, rats were sacrificed. Adhesion formation was not observed in rats grafted with the constructs, whereas severe adhesions were observed in all control rats. Constructs seeded with mesothelial cells isolated from EGFP-transgenic rats clearly revealed that grafted mesothelial cells remained at the host tissue site even after fibrin scaffold degradation. These cells developed more abundant microvilli in vivo than those in vitro. These results show that cultured mesothelial cell sheets are effective in preventing adhesion formation and should reduce postoperative complications caused by adhesion formation.     

Uptake of tritiated leucine by axotomized cervical motoneurons: An autoradiographic study

Kittens, aged 7–9 weeks, were killed in groups of four at survival periods of 0 (unoperated controls), 1, 2, 3, 5, 10, 15, and 28 days following left brachial plexotomy. [³H]Leucine was administered intraperitoneally, 10 μCi/g body wt, 0.25, 0.5, 2, or 16 hr before sacrifice. Qualitative and quantitative histologic and autoradiographic observations were made. On the axotomized side, chromatolysis was qualitatively evident in 97% of C-8 motoneurons 10 days postoperatively. Cellular enlargement was highly variable. Nuclear atrophy was frequent 10–15 days after surgery. Regenerating axons penetrated the neuroma 5 days after plexotomy and reached the ulnar nerve at the level of the olecranon at 28 days and neuromuscular junctions of flexor carpi ulnaris at 60 days. Axotomized cervical motoneurons from kittens sacrificed 0.25 hr after administration of [³H]leucine on the 10th or 15th postoperative day exhibited a relative decrease in the uptake of labeled amino acid. A similar decrease occurred in the 28-day survival sacrificed 0.50 hr after injection of labeled amino acid. In contrast when animals were killed at the 2.0 hr postinjection period, a relative increase in the somal accumulation of tritiated leucine occurred 2 through 28 days post-operatively. A comparable increase obtained 16.0 hr after isotope injection and 2–10 days after plexotomy. The differences in the direction of change observed at short (0.25 and 0.5 hr) and long (2.0 and 16.0 hr) labeling intervals suggest that alterations in accumulation of radiolabeled amino acid by axotomized somas may be related to changes in the types of proteins synthesized or to alterations in the rates of export and/or turnover of cytoplasmic proteins. The increased cytoplasmic radioactivity observed after 2.0- and 16.0-hr labeling intervals preceded light microscopic evidence of axon regeneration and was not dependent on reestablishment of peripheral contacts.     

Division of small intensely fluorescent cells in neonatal rat superior cervical ganglion is inhibited by glucocorticoids

Postnatal genesis of small, intensely fluorescent cells was studied in the rat superior cervical ganglion by combining immunocytochemistry of tyrosine hydroxylase with tritiated thymidine auto-radiography. After injection of tritiated thymidine during the first postnatal week, silver grains were observed over the nuclei of many small cells with intense staining for tyrosine hydroxylase, suggesting that SIF cells are dividing postnatally. Cell counts in ganglia of rats sacrificed 2 h after tritiated thymidine showed that the rate of SIF cell proliferation was highest during the first postnatal week with approximately 20% of SIF cells dividing and that the rate declined thereafter. Counts of labeled SIF cells at 30 days in rats injected with tritiated thymidine on days 0, 2, 4, 6, 8, 10 or 14 revealed a peak of SIF cell birthdays on day 8. In these long-survival experiments, many labeled SIF cells were present in adult superior cervical ganglions. In contrast, only one labeled principal neuron was observed in a total of 450 sections. Glucocorticoid treatment of the rats during the first postnatal week paradoxically increased the number of SIF cells, but inhibited the rate of SIF cell proliferation. Dividing SIF cells immunoreactive for both tyrosine hydroxylase and phenylethanolamineN-methyltransferase were observed in glucocorticoid-treated rats.

These observations suggest that many SIF cells are dividing during the first postnatal week. After cessation of division, these cells either remain SIF cells or die, but do not differentiate into principal neurons. Since glucocorticoids do not stimulate SIF cell proliferation, they must increase the number of SIF cells by biasing the differentiation of precursor cells in the superior cervical ganglion and/or enhancing SIF cell survival.     

Transformation of primary cultured and co-cultured adult rat liver cells by 3'-methyl-4-dimethylaminoazobenzene

Under various conditions of culture and carcinogen treatment, the transformation of liver cells by 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB) was studied. Primary liver cell (PLC) cultures from adult male rats and co-cultures with PLCs of ARL-D8 cells of a liver epithelial-like clear cell line from adult female rats were treated with 0.24 mM 3'-Me-DAB for 6 days. Four of 8 carcinogen-treated PLC cultures contained cells with marker chromosomes, and 3 of the 8 cultures contained gamma-glutamyltranspeptidase (GGT)-positive cells. Three of 5 carcinogen-treated co-cultures contained cells with marker chromosomes, and 2 of the 5 co-cultures contained GGT-positive cells. Pure cultures of ARL-D8 cells were treated for 6 or 12 days with 3'-Me-DAB (0.24 mM)-containing-medium perfused through the liver of adult male rats in situ. In the 6-day treatment, none of 5 carcinogen-treated cultures showed chromosomal abnormality or cytochemically exhibited GGT activity. However, in the 12-day treatment, 2 of the 5 carcinogen-treated cultures contained cells with marker chromosomes, and 2 of the 5 cultures contained GGT-positive cells. None of the control cultures exhibited chromosomal abnormality or GGT-positive cells. In summary, transformation markers increased in ARL-D8 cells when they were co-cultured with PLCs.     

Glycogen changes in the liver cells of young rats following isoprenaline treatment. (Histochemical and ultrastructural investigations).

Glycogen changes were investigated by light and electron microscopy in the liver cells of newborn rats following isoprenaline (IPR) treatment either for 1 day (2 intraperitoneal injections at 12 h interval), or for 8 days (2 intraperitoneal injections daily). On the day following the interruption of both treatments glycogen depletion was observed as compared to control rats, as evaluated by PAS reaction and confirmed by higher total phosphorylase activity. During this stage electron microscopy revealed mainly alpha-particles of glycogen associated to highly dilated smooth endoplasmic reticulum (SER) after 1-day-treatment, and to mostly increased SER after prolonged treatment. In the animals submitted to prolonged IPR administration and sacrificed at later times, glycogen masses intensely PAS-positive were strongly increased, while the activity of total phosphorylase proved accordingly lower than in the rats sacrificed at earlier times. Electron microscopy examination confirmed an increased amount of glycogen (alpha- and beta-particles) differently distributed: beta-particles were more numerous in the liver of the rats sacrificed on the last day of the experiment. At this time SER didn't appear modified as compared to control rats.     

A preliminary study of the use of human adipose tissue-derived stem cells for the treatment of streptozotocin-induced diabetes mellitus in a rat model

The purpose of this study is to determine if intraventricular injection of human adipose tissue-derived stem cells (hADSC) are effective in treating streptozotocin (STZ)-induced diabetes mellitus (DM) in nude rats. Twenty-two adult, male nude rats (strain Crl:NIH-Fox1^RNU) were used to induce diabetes using streptozotocin. A single, 150 mg/kg STZ was injected intraperitoneally. Severity of the induced diabetic state was assessed by daily monitoring of body weight, clinical signs, and blood glucose levels. C-peptide was assessed before ADSC injection (T0) and at 3, 5, and 21 days after ADSC injection. Eight rats (40%) developed DM within 24 h after STZ injection. Of the eight rats that developed DM, five were given 2 million freshly prepared ADSC intraventricularly under echocardiography guidance 10 days after STZ injection and three were only given sterile saline for comparison. Surviving rats were humanly sacrificed 21 days after ADSC injection. The average weight of diabetic rats decreased significantly after STZ injection. ADSC injection had no effect on the body weight of rats. Non-fasting serum glucose levels increased significantly in both groups. In diabetic rats, C-peptide decreased significantly before ADSC injection and seemed to return to normal 21 days after ADSC administration. Results of this preliminary study might suggest a beneficial effect of using hADSC for the treatment of STZ-induced diabetes in adult nude rats.     

Random X-inactivation in a girl with duplication Xp11.21-p21.3: report of a patient and review of the literature.

We describe a 10-month-old girl with abnormal clinical findings and Xp duplication. She showed poor weight gain and developmental retardation, and had several minor anomalies including pigmentary dysplasia (hypomelanosis of Ito). She had a partial short arm duplication in the paternally derived X chromosome, 46,X,dup(X)(p11. 21p21.3), with the normal and duplicated X chromosomes randomly inactivated. These findings indicate that gross functional imbalance in the cells with an active dup(X) chromosome has caused global developmental defects in the patient, and that functional chromosomal mosaicism with respect to the duplicated Xp region has resulted in pigmentary dysplasia. Literature review of 52 patients with partial X duplications revealed (1) random or skewed but not completely selective X-inactivation in 9 of 45 patients examined for the X-inactivation pattern, independently of the size or location of duplicated segments, (2) apparently normal phenotype in 6 of 9 patients with random or skewed X-inactivation, and (3) an abnormal phenotype in 13 of 35 patients with completely selective inactivation of dup(X) chromosomes.     

Effects of PEG-PLA-nano artificial cells containing hemoglobin on kidney function and renal histology in rats

This study is to investigate the long-term effects of PEG-PLA nano artificial cells containing hemoglobin (NanoRBC) on renal function and renal histology after 1/3 blood volume top loading in rats. The experimental rats received one of the following infusions: NanoRBC in Ringer lactate, Ringer lactate, stroma-free hemoglobin (SFHB), polyhemoglobin (PolyHb), autologous rat whole blood (rat RBC). Blood samples were taken before infusions and on days 1, 7 and 21 after infusions for biochemistry analysis. Rats were sacrificed on day 21 after infusions and kidneys were excised for histology examination. Infusion of SFHB induced significant decrease in renal function damage evidenced by elevated serum urea, creatinine and uric acid throughout the 21 days. Kidney histology in SFHb infusion group revealed focal tubular necrosis and intraluminal cellular debris in the proximal tubules, whereas the glomeruli were not observed damaged. In all the other groups, NanoRBC, PolyHb, Ringer lactate and rat RBC, there were no abnormalities in renal biochemistry or histology. In conclusion, injection of NanoRBC did not have adverse effects on renal function nor renal histology.     

Plasma hyperosmolality at the onset of drinking during starvation induced hypovolemia

Elevations of 2–4% in plasma osmolality were noted at the initiation of drinking following 4 days of fasting in rats. Paired animals food deprived for identical time periods but not drinking when blood sampled revealed ad lib levels of osmolality. Drinking initiation by 4 day food deprived rats following access to dry food was also accompanied by 2–4% elevations in plasma osmotic pressure. Substantial decreases in daily water consumption were observed during the 4 day food deprivation period accompanied by a mean plasma volume loss of 32% determined by T-1824 dye injection. Total blood volume loss was estimated to by 27%. Hematocrit was found to be the best estimator of plasma volume in starved rats. The elevations in plasma osmolality did not appear to be the result of uremia due to starvation induced protein catabolism. The data support the position that elevated plasma osmolality accompanies the initiation of drinking during extended fasting while return to ad lib set point osmolality accompanies cessation of drinking.