Lymphocyte-mediated cytotoxicity in vitro. Effect of enhancing antisera.

The ability of antisera to suppress immune responses either in vivo or in vitro is well known. A variety of lymphocyte-target cell systems have been employed to demonstrate inhibition of cell-mediated immunity by antisera in vitro, and skin, tumor, and kidney graft survival have been prolonged by passively administered antiserum in vivo. An in vitro lymphocyte-tumor cell assay system was developed for the purpose of studying the effects of enhancing antisera (in vivo) on lymphocyte-mediated cytotoxicity in vitro. The characteristics of this system with respect to route of immunization, time of harvest of immune cells, lymphocyte:tumor cell ratio, and effect of nonimmune or nonspecifically immune lymphoid cells are presented. Sera capable of enhancement in vivo were tested in this system and shown to inhibit cell-mediated immunity in vitro. Further, in both instances the immunosuppressive effect is mediated by antigen-antibody complexes and not by free antibody alone. Experiments were also carried out to determine the site of action of these suppressive antigen-antibody complexes. Presensitized lymphocytes were exposed to antigen-antibody complexes, washed, and then allowed to interact with fresh tumor cells (not antibody treated). Lymphocytes treated in this manner are incapable of exhibiting cell-mediated immunity in vitro. This evidence supports the concept that the antigen-antibody complexes have a direct immunosuppressive effect on the lymphocyte.

Nuclear fluorescence employing antinucleoside immunoglobulins.

Fluoresceinated antinucleoside globulins were shown to react with the nuclei of L cells. The pattern of nuclear fluorescence was similar to the distribution of nuclear DNA. This reaction was shown to be specific by the following control experiments: 1. Absorption of the specific antibody from an antiadenosine globulin eliminated all fluorescence. 2. Treatment of the cells with nonfluorescent antiadenosine globulin, followed by staining with the fluorescent antiadenosine eliminated almost all of the fluorescence of the nucleus. 3. Treatment of the cells with DNase destroyed the ability of the nucleus to react with antiuridine fluorescent antibodies. 4. Fluoresceinated anti-BSA did not produce nuclear fluorescence. Nuclear fluorescence occurred only in cells harvested during the period of maximum DNA synthesis as measured by the uptake of thymidine. This correlates with the previously demonstrated specificity of the antibodies for denatured DNA.

Renal transplantation between HL-A identical donor-recipient pairs. Functional and morphological evaluation.

16 patients underwent renal transplantation from a sibling donor who was prospectively determined to be ABO compatible and HL-A identical with the recipient. Unidirectional mixed leukocyte reactions were performed; in each instance, lymphocyte stimulation in either direction was not observed. The plasma creatinine 10-68 months after transplantation in these 16 patients ranged between 0.9 and 1.9 mg/100 ml. The creatinine clearance ranged from 48 to 113 ml/min, and the blood urea nitrogen (BUN) ranged between 12 and 35 mg/100 ml. Urine protein excretion varied from 0.11 to 1.86 g/day. Six patients exhibited no detectable clinical episodes of acute rejection; they were treated with azathioprine alone and each of them demonstrated normal or near normal renal histology when biopsy specimens were obtained more than 6 months after transplantation. Nine patients experienced acute rejection episodes that required the use of steroid therapy. The severity of these rejection episodes was variable; they included a mild reduction in renal function with an immediate steroid-induced restoration of function and eventual discontinuance of steroid therapy to severe reduction in function requiring prolonged and moderate doses of steroids without return to normal renal function. Renal histological observations in this group ranged from mild to marked cellular and structural changes which fit the criteria of the rejection. One patient demonstrated a gradual loss of renal function with heavy proteinuria. Biopsy of this allograft demonstrated the recurrence of original disease, i.e., lobular glomerulonephritis. The marked variability in the clinical course and allograft morphology in these 16 patients could be explained by antigenic differences at non-HL-A loci. The presence of minor histocompatibility loci has been well documented in other mammalian species and they are most certainly present in man. The need for their identification and definition is stressed.

Piroxicam-induced renal disease.

Two cases of nephrotoxicity associated with piroxicam therapy are reported. In the first case, severe reversible azotemia with hyperkalemia resolved after discontinuation of the medication. In the second case, biopsy-proved acute interstitial nephritis with immune complex glomerulonephritis and hepatitis improved after steroid therapy. The mechanism for such toxicity may be altered hemodynamics, cellular immunity, or humoral immunity related to prostaglandin inhibition.

Perspective of NIRS measurements early post mortem for prediction of pork quality.

The potential of near-infrared spectroscopy (NIRS) measurements early post mortem was investigated to predict ultimate drip loss, colour, tenderness and intra-muscular fat of pork. Three locations (M. longissimus thoracis, M. longissimus lumborum and M. semimembranosus) in 102 pig carcasses were tested at the end of the slaughter line. A priori variation in pork quality was introduced using an experimental design covering: genotype, lairage time, pre-slaughter handling and day of slaughter. At 1 h post mortem a diode array VIS/NIR instrument (Zeiss MCS 511/522, 380-1700 nm) equipped with a surface fibre optic probe was used and at 1 day post mortem ultimate pH, drip loss, colour and shear force was measured on similar locations. Results indicated that it was possible to predict intra-muscular fat content (correlation (R(2) of 0.35 with multiple linear regression), standard error of prediction (SEP)=3.6 g/kg), but the configuration has to be refined for on-line application (bigger aperture). For drip loss no correlation was achieved with the PLS method. Even extremes (low drip loss (<2.5%) or high drip loss (>4.5%)) in drip loss were not discriminated. Predicting drip loss with NIRS early post mortem is not successful, although NIRS in the slaughter line has potential as a fast predictor of intra-muscular fat. Possibilities for using the NIRS technique to get to know more about muscle metabolism and post mortem changes are promising.

Aggression, suicide, and serotonin: relationships to CSF amine metabolites.

In an earlier, separate study, the authors found that human aggression and suicide (a specific aggression-related behavior) were associated with lower levels of CSF 5-hydroxyindoleacetic acid (5-HIAA), a serotonin metabolite. That study focused on subjects with personality disorders without affective illness. In the present study they examine the life history of aggression and history of suicidal behavior in 12 subjects with borderline personality disorders without major affective disorder. Histories of aggressive behaviors and of suicide attempts were significantly associated with each other, and each was significantly associated with lower 5-HIAA levels. Altered serotonin metabolism may be a highly significant contributing factor to these behaviors in whatever diagnostic group they occur.

Rapid chilling cannot prevent inferior pork quality caused by high preslaughter stress.

The present experiment investigated whether increasing chilling rate could improve meat quality in pigs exposed to either minimal or high stress immediately preslaughter. Pigs (n = 192) were offspring of halothane-free lines. On various days, four groups of 48 pigs were processed at a commercial plant. Within each group, half the pigs were exposed to either minimal or high preslaughter stress. Before entering the cooler at 45 min postmortem, carcasses of both minimal and high preslaughter stress treatments were allocated randomly to either conventional (+4 degrees C for 22 h) or rapid (three-phase chilling tunnel: -15, -10, and -1 degrees C for 15, 38, and 38 min, respectively, followed by storage at 4 degrees C until 22 h postmortem) chilling. Temperature and pH were measured in the blood at exsanguination and in the longissimus lumborum (LL) and semimembranosus (SM) muscle at 0.5, 2.5, 4.5, 6.5, and 22 h postmortem. Meat quality attributes (water-holding capacity and objective color measurements) were assessed on the LL. Preslaughter stress level affected pH and temperature in both blood and muscle, with lower (P < 0.001) pH values and higher (P < 0.001) temperatures for pigs exposed to high vs. minimal stress. Rapid chilling led to a faster (P < 0.001) temperature decline regardless of preslaughter stress level. Rapid chilling did not (P > 0.05) influence the rate of pH decline in the LL muscle, but reduced (P = 0.061) pH decline in the SM. Rapid chilling, as opposed to conventional chilling, decreased (P < 0.05) electrical conductivity in the LL, regardless of preslaughter stress; however, it could not compensate for the detrimental effect (P < 0.05) of stress on drip loss, filter paper moisture absorption, and meat color (L* value). Results from the present study indicated that increasing chilling rate is not a suitable method to resolve pork quality problems caused by inadequate preslaughter handling.

Development of a polymer endovascular prosthesis and its implantation in porcine arteries.

A polyethylene-terephthalate braided mesh stent has been developed for application in the (coronary) arterial tree. In vitro measurements showed that the radial pressure delivered by this device was in the same range as that of a stainless steel stent. Hysteresis-like behavior, however, occurred after constraining the polyester stent for a period of only 15 minutes on a delivery system for percutaneous implantation. This implies that the polymer stent must be mounted on this delivery system immediately before the placement procedure, and that either a diameter in the unconstrained condition must be selected, which is considerably larger than the diameter of the target vessel, or stent expansion has to be enhanced by balloon expansion. Taking into account the results obtained during the in vitro studies, we investigated the angiographic patency and histologic features after implantation of this polyester stent in peripheral arteries of pigs. In four animals eight stents were placed. Except for heparin during the implantation procedure only, antithrombotic or antiplatelet drugs were not administered. After 4 weeks repeat angiography was performed. Angiography revealed that five of the six correctly placed stents were patent. At autopsy, two additional patent stents proved to be located in the aortic bifurcation, probably due to failure of the delivery system. Quantitative assessment showed that the mean luminal diameters of the site of stent placement were 3.3 +/- 0.2 mm before, 3.2 +/- 0.2 mm immediately after, and 3.1 +/- 0.3 mm at 4 weeks after implantation. Histology demonstrated an inflammatory reaction of variable severity around the stent fibers. Quantitative histologic measurements showed that the thickness of the neointima was 114 +/- 38 mum after 4 weeks. In conclusion, polyester stents can be constructed with mechanical properties similar to stainless steel stents. Hysteresis-like behavior of polyester stents, however, influences the selection of the nominal stent diameter as well as the forces exerted to the vessel wall. After implantation in porcine peripheral arteries, five of six correctly placed stents were patent at 4 weeks. The extent of neointimal proliferation was similar to that observed after placement of metal stents in swine, despite the presence of a more pronounced inflammatory reaction.