Two major autoantigen-antibody systems of the mitotic spindle apparatus

OBJECTIVE: To characterize human autoantigen-antibody systems related to the mitotic poles and spindles. METHODS: Thirty-seven human sera with autoantibodies staining mitotic poles and spindles in indirect immunofluorescence (IIF) studies were further characterized by immunofluorescence on mitotic cells and by immunoblotting and immunoprecipitation. Clinical diagnoses meeting the American College of Rheumatology criteria were based on chart review and interview with the corresponding physicians. RESULTS: Two autoantibody systems reactive with mitotic poles and spindles were defined. Type 1 nuclear mitotic apparatus (NuMA-1) antibodies were identified in the serum of 30 patients. Interphase cells showed a fine, speckled, nuclear staining, while mitotic cells had bright staining of the rim of the centrosomes and light staining of the spindles proximal to the centrosomes. In telophase, the staining shifted from the centrosomes to the reforming nuclei. On immunoblotting, anti-NuMA-1 sera reacted with a 210-kd protein. The reactivity of these sera was identified (with the aid of reference antibodies) as the previously described NuMA antigen-antibody system. Clinical information was available for only 17 of the 30 patients with anti-NuMA-1; of these, 17 (53%) had clinical and lip biopsy findings that met the criteria for Sj?gren's syndrome. NuMA-2 antibodies were found in the sera of 7 patients. Interphase cells showed no nuclear or cytoplasmic staining, but mitotic cells had brightly stained poles and spindles. At anaphase/telophase, staining shifted to the midbody and the intercellular bridge. Anti-NuMA-2 sera immunoprecipitated a protein of 116 kd. This group of patients was more heterogeneous and had both systemic and organ-specific autoimmune diseases. CONCLUSIONS: NuMA protein (here called NuMA-1) and a 116-kd protein (here called NuMA-2) are the major targets of the autoimmune response in the mitotic apparatus, since most of the selected sera (based on IIF staining of the mitotic spindles and poles) recognized 1 of these 2 antigens.     

Performance and metabolic responses of young dairy calves fed diets supplemented with chromium tripicolinate

Forty-two Holstein calves were used to study performance and metabolic responses when milk replacer and then postweaning starter were supplemented with 1 ppm of Cr as Cr-tripicolinate. From birth through 8 wk of age, supplemental Cr tended to improve the growth performance of bull calves but not of heifer calves. Starter intake and feed efficiency were not affected by supplemental Cr. From 1 to 5 wk of age, plasma cortisol concentrations sampled just prior to feeding decreased, and concentrations of insulin-like growth factor-I increased. All calves appeared to become less sensitive to insulin as they aged. From 1 to 5 wk of age, plasma glucose and insulin concentrations gradually diverged for all calves; glucose concentrations decreased, and insulin concentrations increased. In addition, glucose clearance rate, measured by i.v. glucose tolerance tests, was more rapid when calves were 2 wk of age than when calves were 8 wk of age. The glucose clearance rate was greater in heifer calves than in bull calves but was not affected by supplemental Cr. Entry of plasma glucose following an i.v. propionate load was also greater in heifer calves than in bull calves but was not affected by supplemental Cr. Plasma nonesterified fatty acids were lower in calves fed milk replacer or starter supplemented with Cr than in control calves, although this effect diminished as calves aged. This finding was considered to be indirect evidence of enhanced insulin sensitivity in calves fed milk replacer or starter supplemented with Cr. Overall, data suggested that supplemental Cr-tripicolinate had minor effects on the metabolism and growth performance of conventionally managed dairy calves. The most notable effects occurred during the initial few weeks of life.     

Persistence of clones of coagulase-negative staphylococci among premature neonates in neonatal intensive care units: two-center study of bacterial genotyping and patient risk factors

From 1 January 1995 until 1 January 1996, we studied the molecular epidemiology of blood isolates of coagulase-negative staphylococci (CoNS) in the Neonatal Intensive Care Units (NICUs) of the Sophia Children's Hospital (SCH; Rotterdam, The Netherlands) and the Wilhelmina Children's Hospital (WCH; Utrecht, The Netherlands). The main goal of the present study was to detect putatively endemic clones of CoNS persisting in these NICUs. Pulsed-field gel electrophoresis was used to detect the possible presence of endemic clones of clinical significance. In addition, clinical data of patients in the SCH were analyzed retrospectively to identify risk factors for the acquisition of positive blood cultures. In both centers, endemic CoNS clones were persistently present. Thirty-three percent of the bacterial isolates derived from blood cultures in the SCH belonged to a single genotype. In the WCH, 45% of all bacterial strains belonged to a single clone. These clones were clearly different from each other, which implies that site specificity is involved. Interestingly, we observe that the clonal type in the SCH differed significantly from the incidentally occurring strains with respect to both the average pH and partial CO2 pressure of the patient's blood at the time of bacterial culture. We found that the use of intravascular catheters, low gestational age, and a long hospital stay were important risk factors for the development of a putative CoNS infection. When the antibiotic susceptibility of the bacterial isolates was assessed, a clear correlation between the nature of the antibiotics most frequently used as a first line of defense versus the resistance profile was observed. We conclude that the intensive use of antibiotics in an NICU setting with highly susceptible patients causes selection of multiresistant clones of CoNS which subsequently become endemic.     

Aberrant RNA processing in a neurodegenerative disease: the cause for absent EAAT2, a glutamate transporter, in amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease that is characterized by selective upper and lower motor neuron degeneration, the pathogenesis of which is unknown. About 60%-70% of sporadic ALS patients have a 30%-95% loss of the astroglial glutamate transporter EAAT2 (excitatory amino acid transporter 2) protein in motor cortex and spinal cord. Loss of EAAT2 leads to increased extracellular glutamate and excitotoxic neuronal degeneration. Multiple abnormal EAAT2 mRNAs, including intron-retention and exon-skipping, have now been identified from the affected areas of ALS patients. The aberrant mRNAs were highly abundant and were found only in neuropathologically affected areas of ALS patients but not in other brain regions. They were found in 65% of sporadic ALS patients but were not found in nonneurologic disease or other disease controls. They were also detectable in the cerebrospinal fluid (CSF) of living ALS patients, early in the disease. In vitro expression studies suggest that proteins translated from these aberrant mRNAs may undergo rapid degradation and/ or produce a dominant negative effect on normal EAAT2 resulting in loss of protein and activity. These findings suggest that the loss of EAAT2 in ALS is due to aberrant mRNA and that these aberrant mRNAs could result from RNA processing errors. Aberrant RNA processing could be important in the pathophysiology of neurodegenerative disease and in excitotoxicity. The presence of these mRNA species in ALS CSF may have diagnostic utility.     

Comparison of mother-father perceptions of their child's self-management of diabetes

This study compared mothers' and fathers' perceptions of their children's self-management of diabetes. Participants included a subsample of 29 paired parents of children and youth with diabetes, ages 8-14, who attended a diabetes camp in the Midwest. Parents completed a self-report structured questionnaire that included content specific to self-management. Results showed high ratings for child responsibility for self-management, style of care delivery, and mother-father self-efficacy in managing specific tasks of diabetes self-management. Although overall ratings were similar, in the majority of measures mothers' ratings were higher than fathers' ratings. Areas of similarity and significant differences in perceptions are identified with implications for practice stated.     

Evaluation of the pathogenicity and virulence of Yersinia species isolated in Edo and Delta States of Nigeria

Sixteen isolates of four species of Yersinia comprising five of Yersinia enterocolitica and Yersinia fredricksenii, four of Yersinia intermedia and two of Yersinia kristensenii isolated from domestic and wild animals in villages in Edo and Delta States, Nigeria, were evaluated for their pathogenicity using laboratory animal models and virulence characteristics tests which included autoagglutinability, calcium dependency for growth, heat-stable enterotoxin production and conjunctivities in guinea pig eye. Results obtained revealed that Yersinia enterocolitica isolates were enteropathogenic as demonstrated by the production of diarrhoea and eventual recovery from faeces, spleen and liver of the infected animals. Three (60%) (2 serotypes 0:3 and 1 serotype 0:8) of the five Yersinia enterocolitica isolates were lethal to the animals. Other Yersinia isolates (Yersinia kristensenii, Yersinia fredricksenii and Yersinia intermedia) were uniformly non pathogenic to the animals. However, a strain of Yersinia intermedia isolate produced diarrhoea in the inoculated animals and caused lethality in guinea pigs and mice, but was negative for autoagglutination test, calcium dependency, conjunctivities, and positive for heat-stable enterotoxin production. We are of the view that this strain may be another Yersinia intermedia--like bacterium, previously isolated in Nigeria. Results therefore, suggest an emergence of a pathogenic Yersinia intermedia species in this environment.