Tissue transplantation. Quality assurance in the banking and utilization of musculoskeletal allografts.

Tissue allografts are being used more frequently, and, as a consequence, an increasing number of hospitals and physician assistants must face the difficult and perplexing task of evaluating their effectiveness and safety. Quality assurance programs are designed to enhance patient care during the transplantation process and should outline the responsibilities of both the tissue bank and the hospital using the allograft. Recommendations for the establishment of quality assurance programs at hospitals are made based on guidelines established by national certifying agencies. It is of paramount importance to recognize the areas of responsibility and to delineate acceptable standards of practice when developing and implementing quality assurance programs for tissue transplantation.

Argyrophilic nucleolar organizer regions in breast carcinoma. Correlation with DNA flow cytometry, histopathology, and lymph node status.

Argyrophilic nucleolar organizer regions (AgNOR) have been correlated with proliferative activity of neoplasms. Increased AgNOR may reflect increased proliferative activity of cells or ploidy. To explore this hypothesis, 41 breast carcinomas were processed for AgNOR silver staining and DNA flow cytometry. AgNOR counts were expressed as mean AgNOR/nucleus and percentage of tumor cells with more than five AgNOR/nucleus. The first count was designated mean AgNOR or mAgNOR, and the second count was designated AgNOR proliferative index or pAgNOR. Using Mantel-Haensel statistical analysis, carcinomas that exhibited mAgNOR of 2.4 or more had a high likelihood of aneuploidy (P less than 0.0001), an S-phase fraction of more than 5.8% (P less than 0.003), or a diameter greater than 2 cm (P less than 0.007). In addition, tumors with pAgNOR of 8% or more showed a statistically significant correlation with aneuploidy (P less than 0.004), tumor grade (P less than 0.04), and a more significant one with high S-phase fraction (P less than 0.0001). No significant correlation was obtained between pAgNOR and tumor size or lymph node status. These data indicate that AgNOR quantitation reflects changes in DNA ploidy and cell proliferation. They also suggest that the mean AgNOR counts correlate best with the DNA mass or ploidy and that the frequency of cells with higher AgNOR count best reflects proliferative activity or S-phase fraction.

Argyrophilic nucleolar organizer regions in Wilms' tumors and related lesions: a histologic and chromosomal correlation.

The argyrophilic nucleolar organizer regions (AgNORs) have been extensively investigated in different neoplasms, and their increased number has been frequently linked to aggressive tumor behavior. To evaluate this association, 22 specimens from 21 patients with Wilms' tumors and related lesions were stained by the AgNOR silver-staining technique in formalin-fixed, paraffin-embedded tissue. The aim of the study was to assess the significance of AgNOR count independently or in relation to histology in predicting the behavior of these neoplasms. Eleven nonneoplastic pediatric kidneys were used as controls. The controls had a range of 1.55 to 2.26 AgNOR/nucleus with a mean +/- SD of 1.91 +/- 0.22. The tumors showed a range of 1.25 to 2.86 AgNOR/nucleus with a mean +/- SD of 2.10 +/- 0.38 with obvious overlap between tumors and controls. The two unfavorable histology Wilms' tumors and the malignant rhabdoid tumor showed no increase in the number of AgNOR (1.66, 2.04, and 2.05, respectively). We further quantitated the nucleolar organizer regions (NOR) in metaphase spreads of three of the neoplasms, compared those with the patients' peripheral blood, and showed no difference in NOR numbers except for an occasional decrease in G-group NORs in the tumor cells. Tumors that had metaphase NOR counts of 6 to 9/nucleus had a mean interphase AgNOR count of less than 2.5. These results suggest that quantitative analysis of AgNORs in Wilms' tumors and related lesions is not a reliable predictor of aggressive tumor behavior. The discrepancy between metaphase and interphase NOR counts indicates that AgNOR counts are merely the reflection of the spatial arrangement of NOR-bearing chromosomes.

Multiparametric analysis of peripheral blood in the normal pediatric population by flow cytometry.

Lymphocyte subset analysis was performed on 114 healthy children and 84 healthy adults. Samples were prepared by a whole blood lysis technique and analyzed by flow cytometry. The percentage and total number of CD3+, CD4+, CD8+, and CD19+ lymphocytes were calculated for each of six age groups. A direct correlation with age was seen in the percentages of CD3+, CD4+, and CD8+ lymphocytes. The absolute number and percentage of total lymphocytes, the percentage and absolute number of CD19+ lymphocytes, and the absolute number of CD3+ lymphocytes decreased with age. No significant correlation with age was observed for white blood cells, the absolute number of CD4+ and CD8+ lymphocytes, and the CD4+/CD8+ ratio.

Concurrent use of the oxfendazole pulse release bolus and the monensin rumen delivery device in young grazing cattle.

When they were turned out to grass in May 1987 for their first season, 10 calves were dosed with a 5 x 750 mg oxfendazole pulse release bolus (OPRB) and a monensin sodium rumen delivery device (RDD); eight calves received one OPRB; 10 calves received one RDD and eight calves received neither bolus. Each group was set-stocked on individual paddocks which had been grazed during the previous season by cattle which developed clinical parasitic gastroenteritis and bronchitis (husk). In July, before they were due to be moved to new pastures in mid-summer, and before they were dosed strategically with levamisole HCl, some of the calves not dosed with an OPRB succumbed to clinical parasitic gastroenteritis and husk and received emergency anthelmintic treatment, after which no further clinical episodes occurred. The 'dose and move' strategy was implemented in early August after which both groups not dosed with an OPRB were set-stocked together until the trial ended on October 14, 147 days after turn out. The two groups of calves which had received the OPRB were also moved to new pasture and set-stocked together until the end of the trial. No evidence of clinical helminthiases developed in either of the two groups of calves dosed with OPRBs and their faecal worm egg and larval counts, and plasma pepsinogen activities remained low. They gained significantly more weight than the two groups of calves not dosed with OPRBs (P less than 0.001). The bolus types were compatible and induced no untoward side-effects when used together.

Control of naturally acquired bovine parasitic bronchitis and gastroenteritis with an oxfendazole pulse release device.

Four groups, each of six male Friesian calves, were set-stocked on separate 0.66 ha paddocks from May 7 until October 23 1986. Each of the animals in groups 1 and 4 was dosed with an oxfendazole pulse release bolus at turn out whereas the animals in groups 2 and 3 were left untreated. Parasite-free naive tracer calves were introduced into each paddock for a limited period 12 days after turn out and again at the end of the trial. No adverse reactions or clinical signs were observed in either of the groups of calves which received boluses. The development of clinical parasitic gastroenteritis in both the untreated groups necessitated the humane slaughter of two animals and emergency anthelmintic treatment of the remainder. The lower plasma pepsinogen concentrations, and lower faecal egg and larval counts and worm burdens post mortem, together with the absence of clinical signs of parasitic gastroenteritis and bronchitis in the treated calves, confirmed the high efficacy of the bolus treatment.

Mononuclear cell phenotyping in a hemodialysis population.

A study of peripheral blood mononuclear cell phenotyping and HIV testing was performed in 39 chronic hemodialysis patients. The following abnormalities were found in comparison to a control population: 1) a lower percentage of lymphocytes in the white blood cell differential count, 2) a lower percentage of total T cells, 3) a lower percentage and absolute number of suppressor T cells and 4) a lower percentage and absolute number of B cells. Both increasing age and blood transfusions were associated with reduced helper cells.

The ability of diamphenethide to control immature Fasciola hepatica in sheep at a lower than standard dose level.

Groups of eight Welsh Mountain sheep were dosed with diamphenethide at the rate of 70 mg/kg bodyweight at either one, four, six or eight weeks after artificial infection with approximately 300 Fasciola hepatica metacercariae. Comparisons were made with similarly infected but undosed sheep and with sheep which were neither infected nor dosed. The good clearance of flukes up to six weeks of age (above 97 per cent on pooled data) was reflected in the plasma concentrations of the accepted liver damage marker enzymes glutamate dehydrogenase and gamma-glutamyl transpeptidase. Highly significant correlations were demonstrated between the numbers of flukes recovered, the plasma levels of these enzymes and haemoglobin and plasma albumin values. At 70 mg/kg, diamphenethide was shown to be able to control F hepatica populations of up to six weeks of age. The systematic use of diamphenethide at this dose level at intervals of up to six weeks during the period of metacercarial challenge should prevent ovine fascioliasis.

Subpopulations of lymphocytes in maternal peripheral blood during pregnancy.

The fetus can be considered an allograft with up to one-half of its MHC antigens being potentially recognized by the mother as foreign. This study compares expression of OKT3, OKT4, OKT8, Kappa, Lambda and Ia antigens on lymphocytes in the peripheral blood of normal non-pregnant women, normal pregnant women, patients who are chronic spontaneous aborters and pregnant insulin-dependent diabetic women. Monoclonal antibodies and cytofluorometric analyses were used for these determinations. There were no significant differences (P = 0.01) between these groups for T-cell markers. A statistically significant (P = 0.001) increased ratio of cells bearing surface immunoglobulin to those expressing Ia antigen (K&L/Ia) was observed between normal non-pregnant controls and women with a history of chronic spontaneous abortion. It is concluded that T-lymphocytes in the peripheral blood do not demonstrate a phenotypic abnormality that would account for the non-rejection of the fetal allograft; however, women with chronic spontaneous abortion may have abnormal B-cell differentiation or T-cell activation that mediates chronic spontaneous abortion.

Antibodies to sperm-associated antigens detected by solid phase assays.

Solid phase assays using radiolabeled (RIA) or enzyme-linked (ELISA) antibodies are described for the quantitation of antibodies to sperm-associated antigens. Antibodies were detected in serum from rabbits immunized with human sperm emulsified in Freund's complete adjuvant and in the plasmas or sera of vasectomized males and patients with suspected antibody-mediated infertility. The RIA and ELISA assays are comparable to each other in sensitivity and specificity, rapid in execution, and inexpensive.

Anthelmintic activity of oxfendazole in pigs.

Eighty-five pigs were artificially infected with Hyostrongylus rubidus, Oesophagostomum species and Ascaris suum. On days 2, 10, 20 or 51 after infection groups of six were treated with oxfendazole premix in food at various dose rates. Subsequently the pigs were slaughtered for comparative worm counts to be made in treated and control animals. The efficacy of treatment against hyostrongylus worms of increasing age was 68, 57, 99.8 and 100 per cent after a 3 mg per kg dose, 75, 75, 99.8 and 100 per cent after a 4.5 mg per kg dose and 81, 83, 99.8 and 100 per cent after a 6 mg per kg dose. In the case of Oesophagostomum species the corresponding figures were 78, 100, 100 and 100 per cent efficacy after a 3 mg dose, 93, 100, 100 and 100 per cent after a 4.5 mg per kg dose and 91, 99, 100 and 100 per cent after a 6 mg per kg dose. Ascaris infestation established too poorly for significant results to be obtained. It is concluded that a dose rate of 4.5 mg oxfendazole per kg body-weight should give practical control of Hyostrongylus and Oesophagostomum species in pigs.