Restricted diversity of the variable region nucleotide sequences of the heavy and light chains of a human rheumatoid factor

The complete nucleotide sequences of the variable region genes of the heavy and light polypeptide chains of a human monoclonal rheumatoid factor (RF) produced from a human-mouse heterohybridoma were determined. The antibody, designated YES8c, contained V kappa III, J kappa 2, VH1, JH4, and a D gene segment of 9 amino acids. The nucleotide sequences and the deduced amino acid sequences of the light chain variable region were remarkably homologous (97-98%) to previously described RF of the Wa idiotypic family (PAY, GLO, CUR, FLO, and GAR) and to that of a V kappa III germline gene (Humkv325). The YES8c heavy chain variable region gene was most closely related to the VH1 gene of the restricted human fetal repertoire, designated 51p1, and also to 3 rearranged VH1 genes that were recently isolated from patients with chronic lymphocytic leukemia. These results suggest that variable region genes of RFs are highly conserved and that YES8c VH, as well as V kappa, may be identical to heavy and light chains expressed during early B cell development.     

Human vascular smooth muscle cells and endothelial cells lack catalase activity and are susceptible to hydrogen peroxide

⁵Cr release as lytic and cell detachment as nonlytic injury were employed to estimate neutrophil-mediated injury of cultured human vascular smooth muscle cells and endothelial cells. The reagents hydrogen peroxide or hypoxanthine-xanthine oxidase produced dose-dependent killing and nonlytic cell detachment, which were specifically inhibited by catalase but not by superoxide dismutase. The concentration of hydrogen peroxide or xanthine oxidase to induce cell detachment was less than lytic dose, suggesting that cell detachment was a much more sensitive assay of injury. Neutrophil-mediated cell lysis averaged 15% at most and was mostly dependent on hydrogen peroxide, while neutrophil-mediated cell detachment was nearly 100% and its dependency on hydrogen peroxide varied from 46% to 60%. These results suggest that vascular smooth muscle cells and endothelial cells in neutrophil-mediated events are destroyed by a hydrogen peroxide-dependent process, mainly via a nonlytic cell detachment mechanism. There was no striking difference of sensitivity to hydrogen peroxide between vascular smooth muscle cells and endothelial cells. Vascular smooth muscle ceils and endothelial cells contained fairly high concentrations of superoxide dismutase, but not catalase, activity. The sensitivity of these cells to hydrogen peroxide but not to superoxide may arise from the fact that these cells lack intracellular catalase activity. The injury of vascular cells, which constitute important components of blood vessels, may lead to vascular injury and subsequent tissue damage.     

Increased concentrations of secretory leukocyte protease inhibitor in peritoneal fluid of women with endometriosis

Secretory leukocyte protease inhibitor (SLPI) is a potent inhibitor of human leukocyte elastase. We investigated whether SLPI was present in the peritoneal fluid of women with endometriosis and to clarify the role of SLPI in the pathogenesis of endometriosis. Western blot analyses revealed that SLPI protein was detected as a 12 kDa band in peritoneal fluid. The peritoneal fluid concentrations of SLPI, elastase and interleukin-6 were assayed by enzyme-linked immunosorbent assays (ELISA). SLPI concentrations and the SLPI/elastase ratio in the peritoneal fluid of women with endometriosis were higher than in samples from women without endometriosis. There was no significant correlation between concentrations of SLPI and interleukin-6 in the peritoneal fluid. Immunohistochemistry using an anti-SLPI polyclonal antibody revealed positive staining in peritoneal macrophages, but not lymphocytes. The present findings suggest that SLPI found in the peritoneal fluid of patients with endometriosis may contribute to the pathogenesis of endometriosis.     

A new approach using DNA fingerprinting for the determination of androgenesis as a cause of hydatidiform mole

A new method of DNA analysis has been used for the determination of androgenesis as a cause of complete hydatidiform mole. This method, using a minisatellite core probe, requires only a small amount of DNA and detects the restriction fragment length polymorphisms (RFLPs) due to allelic differences in the number of tandem repeats containing the core sequence. Southern blot hybridization showed an individual-specific DNA fingerprint, and each polymorphic band in molar tissues could be identified as being of paternal, but not maternal, origin. Some polymorphic bands of paternal DNA were not observed in molar tissues, indicating that endoreduplication of a normal haploid sperm or fertilization by dispermy to an anuclear oocyte with no effective genome could be the cause of complete hydatidiform mole. This method is sufficiently reliable and rapid that differential diagnosis could be made between complete hydatidiform mole, partial mole and hydropic change.     

Trilineage Response to rhG-CSF with Subsequent Clonal Hematopoiesis in a Patient with Severe Bone Marrow Aplasia

We treated a patient with severe aplastic anemia with long-term administration of recombinant human granulocyte-colony stimulating factor (rhG-CSF). When a trilineage response of hematopoiesis was obtained after the first treatment, a chromosomal change [45XX, -7] was observed in 20 of the 20 metaphases examined. Later, we were able to show a monoclonal X inactivation pattern in the phosphoglycerate kinase (PGK) gene in the peripheral blood polymorphonuclear leukocytes and mononuclear cells, indicating the presence of clonal hematopoiesis regardless of the disappearance of the karyotype abnormality. We suggest that it is important to pay close attention to the appearance of clonal hematopoiesis during the administration of G-CSF to patients with idiopathic severe bone marrow aplasia.     

Effects of sodium selenite on methylmercury embryotoxicity and teratogenicity in mice.

Female IVCS mice were fed food containing methylmercury (MeHg) at 0, 15.9, or 31.9 nmol/g food and drinking water containing sodium selenite at 0, 11.4, or 22.8 nmol/ml for 30 days before gestation and thereafter up to Day 18 of gestation. Groups receiving MeHg had (1) affected estrus cycles, (2) decreased litter size accompanied by increased implantation sites, resorptions, dead embryos, and dead fetuses, (3) increased incidence of cleft palate, and (4) retarded fetal growth. Selenite was not teratogenic and even at the high dose, generally only retarded fetal growth. The low dose of selenite when combined with the high dose of MeHg, increased the incidence of resorptions and implantation sites. Embryolethality was decreased and the incidence of cleft palate was increased in the group receiving the high dose of selenite combined with the high dose of MeHg.     

NOD mice with high incidence of type 1 diabetes are not T lymphocytopenic

An autoimmune pathogenesis has been indicated in insulin-dependent (type 1) diabetes mellitus (IDDM). Previously we reported that non-obese diabetic (NOD) mice as an animal model of spontaneously developing IDDM were immunologically characterized by T lymphocytopenia and impaired cellular immunities. The cumulative incidence of diabetes in the T lymphocytopenic female NOD mice was 10-20% by 24 weeks of age. On the other hand, the incidence of diabetes are 80-90% in the female NOD/Shi-Sendai (S), in whom proportion of lymophocyte subsets has not been known yet. Therefore, we examined the spleen cells of female NOD/Shi-S and female NOD/Shi with high incidence of diabetes, and of female Jcl:ICR as a control. Cell numbers, populations of T cells (Thy 1.2+, Lyt-1+ and Lyt-2+), B cells (surface-Ig+), NK cells (acialo GM1+) and responsiveness to Concanavalin A were analyzed as immunological parameters. In contrast to the T lymphocytopenic NOD, these immunological parameters were not impaired in the NOD/Shi-S and NOD/Shi in comparison to those of Jcl:ICR. The results indicate that there may be a positive association between the incidence of diabetes and T cell number and functions in female NOD mice.     

Recent demographic and injury trends in people served by the Model Spinal Cord Injury Care Systems

OBJECTIVE: To identify trends in the demographic and injury data of persons with spinal cord injury (SCI). DESIGN: Consecutive case series. SETTINGS: Model Spinal Cord Injury Care Systems throughout the United States. PATIENTS: A total of 25,054 persons admitted to a Model Spinal Cord Injury Care System within 365 days of injury between 1973 and 1998. RESULTS: Many trends and changes have been noted in the clinical features of patients who have been admitted to the Model SCI Care Systems. Average age at time of injury is rising; persons older than 60 comprise 11.5% of all persons enrolled in the National Database during the 1994-1998 period. Although the overall male-to-female ratio is greater than 4:1, the proportion of males has decreased significantly in recent years. Violence-related injuries have increased dramatically from 13.9% in 1973-1977 to 21.8% in 1994-1998. Since 1973, the proportions of injuries resulting from vehicular crashes and sports declined while injuries from falls increased. Injury continues to occur most commonly in the summer. When age, race, and gender are considered, violence is a more common cause of injury among individuals who are younger, male, or African American. Complete injuries were more common among younger individuals and among men than among older adults and women. CONCLUSION: Trends in the national database provide valuable data for tracking groups at risk for traumatic SCI.     

Cardiovascular depression and stabilization by central vasopressin in rats

The role of endogenous vasopressin in cardiovascular homeostasis was examined using vasopressin-deficient rats (Brattleboro) (n = 194) and their parent strain, Long-Evans rats (n = 181). Mean arterial pressure (blood pressure) and heart rate were measured every 4 seconds with or without infusion of drug solution for 21 hours, and mean values and their standard deviations (lability) were calculated. Blood pressure in Brattleboro rats (116 +/- 1.1 mm Hg, mean +/- SEM) was significantly higher than that in Long-Evans rats (96 +/- 0.7 mm Hg, p less than 0.001), whereas heart rates (381 +/- 3.3 and 375 +/- 2.9 beats/min, respectively) were similar. The lability of blood pressure and heart rate in Brattleboro rats (9.2 +/- 0.1 mm Hg and 42.3 +/- 0.7 beats/min) was also greater than that in Long-Evans rats (6.7 +/- 0.1 mm Hg, p less than 0.001 and 38.4 +/- 0.8 beats/min, p less than 0.01, respectively). In Brattleboro rats, intravenous vasopressin (0.1 ng/kg/min or 0.6 ng/kg/min) did not affect blood pressure, although it did reduce heart rate and decreased lability of blood pressure and heart rate. Intracerebroventricular (central) infusion of vasopressin (2 pg/kg/min) in Brattleboro rats induced initial hypertension and tachycardia followed by long-lasting hypotension and bradycardia, whereas in Long-Evans rats it induced only hypertension and tachycardia. In both strains, central vasopressin dramatically decreased the lability of blood pressure and heart rate. Neither intravenous (0.2 ng/kg/min) nor central desmopressin (2 pg/kg/min or 0.2 ng/kg/min), a V2 renal receptor agonist, changed any of these parameters in Brattleboro rats, although both diminished urinary volume. Neither intravenous (50 ng/kg/min) nor central (3.3 pg/kg/min) d(CH2)5-Tyr(Me)-arginine vasopressin, a vasopressin V1 receptor antagonist, modulated any of these parameters in Long-Evans rats. These results suggest that endogenous as well as exogenous vasopressin acts centrally as a cardiovascular inhibitor and stabilizer through a receptor mechanism other than V1 or V2 receptor mechanisms.