'Many are called...': participation by minority leaders in an AIDS intervention in San Francisco. Though the AIDS epidemic has posed a significant threat to minority people, especially African Americans and Latinos, minority community response to the epidemic has lagged significantly behind the organized efforts initiated by the European-American gay community. To test the efficacy of an educational intervention in increasing the involvement of minority leaders in AIDS-related efforts, we identified and recruited 166 African-American and Latino leaders to participate in a randomized controlled trial. This report describes the identification and recruitment of leaders to the study. Overall, participation was low. Rates of participation were not associated with race but were significantly associated with category of leadership--that is, whether the individual was a religious leader, an agency director, an elected official, or an informal leader, with religious leaders least likely and agency directors most likely to participate. We discuss the implications of these findings for improving epidemic response in minority communities.


Oxytalan fibres in proliferative vitreoretinopathy. The distribution of elastic and related fibres was studied in 29 fibrocellular membranes associated with proliferative vitreoretinopathy using histochemical, immunohistochemical and ultrastructural techniques. Ten subretinal and ten epiretinal membranes were subjected to light microscopical investigation, whilst the remaining nine membranes were examined by transmission electron microscopy. Oxytalan fibres, an elastic fibre precursor that is found in a variety of normal and pathological connective-tissue matrices, were identified in all of the specimens. Mature elastic fibres were not observed in any of the 29 membranes. The demonstration that oxytalan connective-tissue fibres contribute to the infrastructure of PVR membranes adds further evidence to the suggestion that this proliferative tissue might be the result of a repair mechanism.


Results of extracapsular cataract extraction with intraocular lens implantation in eyes with uveitis and Fuchs' heterochromic iridocyclitis. A total of 63 cataractous eyes of 48 patients (group 1) with uveitis and 12 eyes of 12 patients (group 2) with Fuchs' heterochromic iridocyclitis underwent extracapsular cataract extraction (ECCE) with capsular bag fixation of a posterior-chamber intraocular lens (PC OIL). A perioperative regimen of systemic dexamethasone (0.8-1.5 mg/kg) was initiated. This dose was tapered by 20 mg/day, reaching zero within 4-8 days. Intraoperatively, synechiolysis was required in 51 of 63 eyes in group 1 and a sector iridectomy was necessary in 42 eyes. A fibrinous reaction occurred in 7 eyes. During a mean follow-up period of 2 years, 7 eyes developed uveitis recurrences that were successfully treated with topical and systemic steroids in 4 eyes; the other 3 eyes developed cystoid macular edema. Overall, mean visual acuity improved from 20/120 to 20/50. Intraoperatively, 3 patients in group 2 developed a transient hemorrhage from atypical anterior-chamber-angle vessels. Postoperatively, 3 eyes showed a transient fibrinous reaction. The mean visual acuity improved from 20/240 to 20/30. In patients with Fuchs' heterochromic iridocyclitis, ECCE with PC IOL implantation renders good intermediate results. In other selected patients with uveitis, in-the-bag IOL implantation may be performed with an acceptable risk. As judged from their improved visual acuity, 57 of 63 eyes (90.5%) benefited from the procedure.


Measurement of potential visual acuity in 343 patients with cataracts. A prospective clinical study. In 343 patients with cataracts (the largest consecutive series in which this subject has been addressed) and 63 patients with clear optic media, who served as the control group, potential visual acuity was measured with the Guyton/Minkowski Potential Acuity Meter (PAM). The control group included 32 cases of macular degeneration, 22 cases of glaucoma, and 9 cases of amblyopia. The cataract group comprised 46 patients with macular degeneration, 46 with glaucoma, and 8 with amblyopia; 5-7 days postoperatively, corrected visual acuity was measured in this group. Because of dense cataracts in 59 patients, no PAM value could be obtained. The visual acuity of 70.4% of the patients, in whom complete data were obtained, was within one Snellen line of the predicted value or better (21%). In general, postoperative predictions were accurate in patients with moderate cataracts. The results were not affected by glaucoma or amblyopia. In 17.5% of patients with macular degeneration, false-positive results were obtained. The control group, in which distance visual acuity was compared with the PAM values, rendered comparable results, with 86.9% of the PAM predictions being correct.


Perimetric follow-up in glaucoma with a reduced set of test points. The global mean defect (GM) is probably the most useful visual field index for glaucoma follow-up. We compared 50 regional subsets of test locations to estimate the GM. Using the data on 424 automated fields of 257 patients with either primary open-angle glaucoma or ocular hypertension, we calculated the partial sample error of the regional mean defect as compared with the GM. In many regions, the measured sample error was greater than expected for a representative sample of points. Some sample errors were up to 5 times larger (regions "upper hemifield" and "lower hemifield"). Only a few subsets proved to be representative of the whole field, namely, regions "ring 3" (10 degrees-15 degrees), "ring 4" (15 degrees-20 degrees) and "ring 5" (20 degrees-25 degrees). The use of such programs for follow-up is very accurate for staging. Moreover, theoretical calculations reveal that trend analysis is even more significant if the reduction in examination time is combined with a proportional increase in examination frequency.


The Aulhorn flicker test: possibilities and limits. Its use in optic neuritis for diagnosis, differential diagnosis, monitoring the course of the disease, and assessing the effect of oral prednisolone. The Aulhorn flicker test, which measures subjective brightness in relation to flicker frequency, is of high specificity (98%) and sensitivity (85.5%) in active optic neuritis (ON). The test has proved to be valuable in the diagnosis and differential diagnosis of ON and in monitoring the course of the disease. The first part of this paper gives an overview based on more than 10 years of experience and on the evaluation of more than 1000 eyes, 250 of them with active ON. The possibilities and limits of the test are analyzed. The second part of the paper presents new results of a prospective, randomized, and controlled treatment study. The Aulhorn flicker test was used to determine whether a prednisolone effect would be assessable by means of this subtle parameter. A total of 50 patients were treated orally with either methylprednisolone (100 mg/day initially) or vitamin B1 (100 mg/day), 38 of them in a double-blind fashion. Although a trend toward faster recovery in favor of prednisolone treatment was noted, no long-term benefit was observed after 1 year.


Influence of windshield tint and tilt on recognition distance under mesopic conditions. The influence of the angle of inclination and tint of windshields on recognition distance was tested in 94 subjects aged 20-83 years. Both clear and tinted (heat-absorbing) windshields were tested. Light transmission (in the vertical position) was 90.3% for the clear windshield and 81.0% for the tinted windshield. The windshields could be set at angles of 0 degree (vertical), 30 degrees, 50 degrees and 70 degrees of inclination. The tests were performed at night with low-beam headlights (mesopic conditions). A Landolt ring (diameter 43.5 cm; contrast 1:1.5) served as a test stimulus; it was painted on a disc 87 cm in diameter that could be rotated in steps of 45 degrees. The mean recognition distance without the windshield was 32.2 m +/- 5.5 m (mean +/- SD). It decreased to 31.0 m +/- 5.6 m (29.0 m +/- 5.3 m) by using a clear windshield in the vertical position (values for the tinted windshield are shown in parentheses): 30.1 m +/- 5.6 m (27.9 m +/- 5.3 m) with a 50 degrees angle of tilt, and 28.5 m +/- 5.4 m (26.9 m +/- 5.1 m) with a 70 degrees angle. The differences between the recognition distances obtained using the various tilts and tints were small but statistically significant (P < 0.01 in all cases: Student's paired t-test). Since the performance of the human eye is stressed to its limits during nighttime driving, our study suggests that extreme tilt of the windshield should be avoided and that the option of delivering cars equipped with a heat-absorbing but clear front windshield should be provided.


Pterygium excision using 193-nm excimer laser smoothing and topical mitomycin C. A total of 31 eyes with primary pterygium and 24 eyes with recurrent pterygium underwent surgical treatment and were followed for at least 1 year. The surgery involved a conventional bare-sclera technique combined with excimer laser smoothing of the wound bed. Postoperatively, the eyes were medically treated with topical mitomycin C (0.02%) for 4 days and with steroids for several weeks. No recurrence was noted within the 1st year in the primary pterygium group, whereas recurrence was observed in 3/24 eyes that had been treated for recurrent pterygium, equivalent to a recurrence rate of 12.5% in this subgroup. Nine eyes (16.4%) had gained 2 lines or more in visual acuity by 1 year after surgery. Granuloma developed in 3 eyes, and dellen formation was detected in 1 eye after surgery. The combination of surgical management with local mitomycin therapy may be considered to be a safe and effective technique for the removal of pterygium. Although the visual results were good, the role of excimer laser smoothing in the treatment of this disorder needs further investigation.


Six years' experience with systemic cyclosporin A prophylaxis in high-risk perforating keratoplasty patients. A retrospective study. Starting with single cases in 1985, we have routinely applied systemic cyclosporin A since 1987 in high-risk keratoplasty patients for a period of up to 37 months postoperatively. In all, 69 eyes undergoing 74 perforating keratoplasties have thus far been treated. A considerable percentage of them initially suffered from chemical burns or from endogenous eczema associated with chronic atopic inflammation. All patients were followed closely and their courses were reevaluated retrospectively. Our current conclusions are that (1) immune reactions are efficiently inhibited with constantly effective blood levels of cyclosporin A, which has enormously increased the rate of keratoplasty successful in these patients; (2) if drug blood levels fall too low due to noncompliance of the patient or to metabolic disturbances, immune reactions may occur, especially during the early postoperative months; (3) serious chronic surface problems, which are especially associated with chemical burns, atopic inflammation, and other chronic kerato-conjunctival diseases, are partly ameliorated but not completely eliminated (surface disorders are presently ranked as the number one cause of transplant failure, whereas immune reactions can be effectively suppressed by cyclosporin A); and (4) close medical follow-up has revealed no serious systemic complication of cyclosporin A prophylaxis over periods of up to 37 months. The present study was not conducted in a prospective, double-blind, controlled fashion.


Mantle cell lymphoma: an update.

Mantle cell lymphoma: an update. Mantle cell lymphoma is a distinctive pathologic entity that incorporates the previous histopathologic categories of centrocytic lymphoma and lymphocytic lymphoma of intermediate differentiation. These lymphomas are characterized by common histologic and immunologic characteristics that suggest derivation from the follicular mantle zone. Mantle cell lymphomas are characterized by the t(11;14) (q13;q32) translocation and its molecular counterpart bcl-1 rearrangement. This translocation activates a gene called BCL-1/PRAD-1. The identification of the BCL-1 gene product as a cyclin has added a new dimension to our understanding of the variety of mechanisms involved in lymphomagenesis.


The role of adhesion molecules in multiple myeloma. Human myeloma plasma cells had been considered to express few surface antigens until recently. The past two International Workshops on Leucocyte Differentiation Antigens have shown that myeloma cells can express a range of surface molecules, and it has become clear that many of these have adhesive functions. The identification of ICAM-1 (CD54) and H-CAM (CD44) on human plasma cells was the initial observation, and other antigens such as N-CAM (CD56) and LFA-3 (CD58) have been confirmed as features of malignant plasma cells in particular. The degree of expression of LFA-1 (CD11a) remains to be characterised fully. It seems probable that the loss of some adhesion structures may be associated with increased malignancy and plasma cell leukaemia. At the present time there are few studies relating to the function of these molecules, although homotypic adhesion appears to occur, and it is likely that such studies will shed light on the pathogenesis of myeloma.


t(2;18) and t(18;22) variant chromosomal translocations in B cell malignancies. Variant translocations (2;18 and 18;22) are described in this review. The chromosomal and molecular findings of these translocation of BCL2 and their effect on possible BCL2 gene activation is discussed. Unanswered questions still remain and these include why this is so rare compared to the 25% incidence recorded for translocations in Burkitt's lymphoma. Further studies are obviously still needed in order to determine the true frequency of these findings and their distribution in the various B-cell disorders.


Karyotype instability in myelodysplastic syndromes--a specific step in pathogenesis preceding clonal chromosome anomalies. In a prospective study, 20 (11.6%) out of 172 patients with myelodysplastic syndromes (MDS) and two patients with aplastic anemia showed an increased spontaneous karyotype instability. No clonal chromosomal aberrations were detectable at the time of first examination. In three patients clonal anomalies developed during the course of the disease. In 17 out of 22 cases (77.2%) chromosomes 1, 5 or 7 were involved in nonclonal anomalies. The same chromosomes were frequently affected in patients with clonal abnormalities too. Our data suggest that spontaneous karyotype instability may be specific to a high proportion of cases with MDS and may constitute the source of clonal chromosome anomalies in these disorders.


A pilot study of prophylactic aerosolized amphotericin B in patients at risk for prolonged neutropenia. Invasive aspergillosis continues to be a significant cause of morbidity and mortality in patients with prolonged neutropenia. We performed a phase I trial of escalating doses of aerosolized amphotericin B given by a face mask nebulizer system with a disposable bacterial exhale filter. Five, 10, 15, and 20 mg of drug were dissolved in sterile water and inhaled over 10 to 15 minutes twice daily. Tolerance was studied in 26 patients (18 transplant recipients, and 8 leukemia patients). No side effects were observed at any dose level. Prophylactic treatment ended for 14 patients (54%) when intravenous (IV) amphotericin B was begun empirically for antifungal coverage following fevers. Eleven patients (43%) continued inhaled amphotericin B until blood counts recovered. One patient was taken off study when she developed cardiogenic pulmonary edema. No patient developed clinically suspicious or pathologically documented infection with invasive aspergillosis. Prophylactic aerosolized amphotericin B is well tolerated at 5, 10, 15, and 20 mg twice daily dosing. In addition, prophylactic aerosolized amphotericin B does not appear to sensitize patients to the subsequent use of IV amphotericin B. Although this study suggests that prophylactic inhaled amphotericin B is well tolerated and effective, a large scale controlled trial is needed.


Serum interleukin-6 levels in patients with thrombocytosis. Interleukin-6 (IL-6) has been shown to increase platelet counts in several animal models and to enhance megakaryocytopoiesis in vitro. In order to investigate the possible relationship between IL-6 and thrombocytosis, serum IL-6 levels in patients with platelet counts > or = 6 x 10(5)/microliters were measured using an IL-6-responsive bioassay. A cohort of healthy volunteers with normal platelet counts was used to establish a control mean serum IL-6 level [2.19 U/ml +/- 1.08 SD (range 0-5.5)]. Patients with primary thrombocytosis had a mean serum IL-6 level not significantly different from controls. In comparison, serum IL-6 levels of patients with reactive thrombocytosis were significantly greater than controls (38.3 U/ml +/- 94.6; range 0-933; P < 0.001). Although no significant correlation was observed between the degree of serum IL-6 elevation and the height of the platelet count in any individual, elevated serum IL-6 was highly correlated with reactive thrombocytosis.


Detection and characterization of circulating immune complexes in peptic ulcer. Immunological factors have been implicated in the pathogenesis of peptic ulcer following the demonstration of autoantibodies against IgA in patients with this disease. We investigated whether circulating immune complexes were present in this condition, and what type of immunoglobulins were involved. The sera of 37 patients with duodenal ulcer and 8 with gastric ulcer were tested for the presence of circulating immune complexes using the polyethylene glycol (PEG) assay and the results compared with those in 79 controls. The precipitate was dissociated and the levels of IgA, IgG and IgM estimated using the single radial immunodiffusion technique. Simultaneous estimation of these immunoglobulins in the serum was also done. Autoantibodies against IgA in the serum were tested using the ELISA test. Eleven patients (6 with duodenal ulcer and 5 with gastric ulcer) were found to have circulating immune complexes. The mean protein content of the PEG precipitates was significantly higher in patients than in control subjects (p < 0.001). The mean values of all the immunoglobulin isotypes were higher in patients than in controls. The IgA content in the PEG precipitates of positive cases was higher than that in control subjects (p < 0.05). Patients with peptic ulcer have circulating immune complexes which may interfere with normal immunoregulation.


Hip instability in newborns in an urban community. It is important to recognize congenital dislocation of the hip early so that it can be treated effectively and secondary changes prevented. There is little data on the incidence of this condition in India and we, therefore, studied this problem in a large maternity and paediatric centre in New Delhi. Six thousand and twenty-nine consecutive newborns were examined for neonatal hip instability by an orthopaedic surgeon and a paediatrician. Clinical examination was supplemented by X-ray and ultrasonographic examination. One hundred and forty-eight hips in one hundred and thirteen newborns (18.7 per 1000 live-births) were found to be unstable at first examination. Of these 12.1 per 1000 live-births had subluxatable hips and 0.82 per 1000 had dislocated hips. Those affected were more likely to be females, have involvement of the left hip and have had a breech presentation. One hundred and twenty-eight hips in 97 children had stabilized within 2 weeks, although no definite progress to stability was observed in subluxatable, dislocatable or dislocated hips. Two children who had other congenital abnormalities initially required operation, but the hips in all the other children stabilized within 20 weeks of birth with conservative treatment. Neonatal hip instability is common in New Delhi and a screening programme should be initiated to detect its incidence in other parts of India.


Hepatitis B virus infection in hospital personnel. Healthcare workers are known to be prone to infection with the hepatitis B virus. We did this study, as there are only a few reports from India on the prevalence of this infection among hospital personnel, so that a rational and effective immunization policy could be evolved. Eight hundred and sixty-three hospital employees including doctors, nurses, technicians, office workers, orderlies and other ancillary staff were screened by ELISA for both hepatitis B surface antigen and its antibody as indicators of prior infection. Eighty-six (10%) were found to be seropositive for hepatitis B surface antigen alone while in 138 (16%) only the antibody was detected. We did not come across any subject in whom both the antigen and the antibody were present simultaneously. Seropositivity was associated with a history of contact with blood or blood products, accidental parenteral exposure and direct patient contact. A history of close contact with a known patient with hepatitis and a past history of jaundice showed a weaker association, while a history of surgery or dental treatment was not significantly associated with seropositivity. There was a significantly (p < 0.5) higher seropositivity in males than in females. The seropositivity was also related to the employee's socio-economic status. Working in a dialysis unit carried the greatest risk. Energetic steps should be taken in all hospitals for the prevention of hepatitis B virus infection among healthcare workers.


F-SSCP: fluorescence-based polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analysis. A fluorescence-based method for polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analysis, F-SSCP, was developed in which the target sequence is amplified by the PCR using fluorescent primers. The amplified products are then heat-denatured and applied to a water-jacket controlled gel in an automated DNA sequencer. The separated strands are detected as laser-excited fluorescence at the bottom of the gel, and mutations are detected as shifts in the position of the peaks in the fluorogram. The system does not involve radioactivity, and the conditions of electrophoresis are more strictly controlled than in the previous system, which relied on ambient air-cooling to maintain the gel at a constant temperature. The nature of the output data allows direct quantitative interpretation, and so the relative abundance of each allele in a mixture of two or more alleles can easily be estimated. The application of F-SSCP for detection of mutations and loss of heterozygosities of p53 in tumor tissues is reported.


Statistical measures of clinical agreement. Disagreements over clinical findings, diagnoses and management decisions are unavoidable in medicine. It is, however, necessary to ascertain whether they agree 'sufficiently'. Such investigations are often analysed inappropriately using Pearson's product-moment correlation coefficient. In this article, we discuss these as well as other methods, such as the kappa statistic, intra-class correlation coefficient and the graphical procedure to measure agreement using clinical examples.


Mismatch amplification mutation assay (MAMA): application to the c-H-ras gene. We have found that under appropriate conditions, an allele-specific polymerase chain reaction (PCR) can achieve a sensitivity suitable for measuring specific, infrequent mutations in single cell systems or in animal tissues. Using the 12th codon GC-to-AT mutation in the rat c-Ha-ras gene as a model system, we have defined conditions that allow for measurement of mutations present at frequencies as low as one in 10(5) gene copies. Our approach involved the use of PCR primers that created a single mismatch with the mutated allele (GAA) but created a double mismatch with the wild-type allele (GGA). Five out of the six such double-mismatch primers we tested permitted amplification of the mutant allele (GAA) with a high degree of specificity. The specificity of the assay was further enhanced by using a two-step PCR cycle consisting of a denaturation step (1 min incubation at 94 degrees C) and an annealing/extension step (1 min incubation at 50 degrees C) in the presence of 10% (vol/vol) glycerol. Reconstruction experiments using genomic DNA demonstrate that this procedure cna measure the presence of 30 copies of the transforming ras allele present amongst 3 x 10(6) copies of the wild-type allele.


Randomization of genes by PCR mutagenesis. A modified polymerase chain reaction (PCR) was developed to introduce random point mutations into cloned genes. The modifications were made to decrease the fidelity of Taq polymerase during DNA synthesis without significantly decreasing the level of amplification achieved in the PCR. The resulting PCR products can be cloned to produce random mutant libraries or transcribed directly if a T7 promoter is incorporated within the appropriate PCR primer. We used this method to mutagenize the gene that encodes the Tetrahymena ribozyme with a mutation rate of 0.66% +/- 0.13% (95% C.I.) per position per PCR, as determined by sequence analysis. There are no strong preferneces with respect to the type of base substituion. The number of mutations per DNA sequence follows a Poisson distribution and the mutations are randomly distributed throughout the amplified sequence.


Automated DNA profiling by fluorescent labeling of PCR products. DNA profiling has been automated by the fluorescent tagging of amplified variable number tandem repeat (VNTR) loci. This was achieved by the use of fluorescently labeled primers in the amplification of 10 ng of genomic DNA, coupled with laser detection of the products during electrophoresis. The PCR products are sized by co-electrophoresing a standard size ladder mixed with every sample, thereby eliminating errors in size estimation caused by lane-to-lane differences in migration rate. This increases the precision of VNTR characterization and enables alleles that differ by a single 15-bp repeat to be resolved. The system is capable of high throughput: Twenty-four samples are electrophoresed and analyzed within 6 hr. Also, because four different dyes are available, three different loci can be simultaneously characterized with the fourth dye used for the internal standard. Approximately 100 unrelated British caucasians were analyzed at the loci D1S80, D17S5, and ApoB. The probabilities of two unrelated individuals matching by chance (pM) at these three loci were determined to be 0.065, 0.040, and 0.069, respectively, with a combined pM of 1.8 x 10(-4).


The use of hybrid-selected template increases the specificity of the polymerase chain reaction. An efficient method for generating full-length DNA sequences from apparently unsuccessful polymerase chain reactions (PCR) has been developed. In cases where nonspecific background interferes with detection of the PCR product, a second amplification is performed using a nested set of primers. The internal fragment of DNA amplified in this reaction is then blotted to a membrane and used to hybrid-select the desired DNA from the initial amplification. This DNA is eluted and used as the template for a third round of PCR. The re-use of the original primers from the initial reaction enables the final PCR to generate full-length DNA. This technique was used to clone a full-length gene segment 8 from a mutant influenza A/WSN/33 (H1N1) virus after initial PCR attempts had failed.


PCR regimen for enhanced specificity and yield of targeted genomic DNA sequences: ras and p53. By weighting the PCR reaction in favor of specificity for the target sequence in the beginning cycles and for continued efficient amplification of the sequence into later cycles, we were able to show an improvement in the specificity and quantity of amplified ras and p53 sequences. Increased purity and yield of specific products favorably enhanced post-PCR evaluation and interpretation of results using direct sequencing and single-stranded conformation polymorphism (SSCP) analysis when point mutations were present in DNA from tumor cell lines and tissues.


A method for nested PCR with single closed reaction tubes. Toward the goal of reducing diagnostic false positives while retaining high sensitivity, a closed-tube nested PCR procedure has been developed for detecting low-copy-number human immunodeficiency virus (HIV) gag target DNA sequences. Master mix for amplification 2, in a hanging gel matrix at the reaction tube top, remains sequestered from the reaction space of the tube during amplification 1. A severalfold excess of inner over outer primers is built into the procedure to assure the high sensitivity of nested PCR. The master mix for amplification 2 is then introduced into the reaction space by centrifugation, and the second amplification is performed as usual. The closed-tube nested procedure shows sensitivity approaching that of the open-tube control procedure, which detects a single copy of HIV gag target DNA at near-theoretical frequency, typically with microgram yields of specific amplification product.


Rapid synthesis of DNA deletion constructs for mRNA quantitation: analysis of astrocyte mRNAs. A rapid method for the synthesis of DNA fragments for competitive PCR analysis is described. This procedure takes advantage of the fact that if PCR is carried out with a mixture of re-ligated restriction digestion fragments, only those fragments containing binding sites for both PCR primers will be amplified. Following electrophoresis of the amplified mixture, the DNA fragment of desired size can be excised from an agarose gel, reamplified, and used for subsequent competitive PCR. We have used this procedure to synthesize deletion constructs for the rat glial fibrillary acidic protein (GFAP) gene, and have used competitive PCR to determine the levels of this mRNA in primary cultures of rat brain astrocytes.


The effect of a combined purging with mafosfamide and hyperthermia on murine haemopoietic stem cells and leukaemogenic cells. The in vitro purging effect of mafosfamide combined with hyperthermia was studied in a murine model. The survival of normal clonogenic progenitors (d-9 CFU-S and CFU-GM) and WEHI 3-B leukaemic clonogenic cells (CFU-L) were compared. At 37 degrees C, CFU-L proved to be significantly more sensitive to mafosfamide than either of the normal progenitors. When mafosfamide was combined with 42.5 degrees C hyperthermia for 1 h, an additive effect was observed: at a dose of 5 micrograms/ml mafosfamide, the survival of CFU-L was nearly two logs lower than that observed at 37 degrees C, while 37.7% of CFU-S survived the purging. The repopulating capacity of surviving bone marrow CFU-S was not altered: a similar 60 d survival of supralethally irradiated recipients transplanted with comparable graft sizes from purged or non-purged bone marrow was observed. When bone marrow suspensions containing WEHI 3-B cells were purged with 5 micrograms/ml mafosfamide at 42.5 degrees C and the minimal amount of bone marrow cells needed to protect supralethally irradiated mice were injected, leukaemia incidence was reduced to less than 10% as opposed to 100% of those injected with untreated bone marrow. Our results suggest that ex vivo hyperthermia may enhance the purging efficiency of mafosfamide.


A method for clinical purging of myeloma bone marrow using peanut agglutinin as an anti-plasma cell agent, in combination with CD19 monoclonal antibody. Previous studies have shown that the lectin peanut agglutinin (PNA) binds bone marrow plasma cells in the majority of patients with myeloma and does not bind to normal haemopoietic progenitors. This lectin has been used in combination with anti-CD19 monoclonal antibody (moAb) in a system for purging myeloma bone marrow. This has now been scaled up for application to ex vivo treatment of large volumes of bone marrow suitable for autologous bone marrow transplantation. Four bone marrow harvests from patients with myeloma containing 9.5 +/- 4.9% plasma cells were depleted of erythrocytes and mature granulocytes by Ficoll separation using the Haemonetics V50 cell separator. The mononuclear fraction was then purged with magnetic beads coated with PNA and anti-CD19 moAb. The system proved highly efficient with removal of all detectable plasma cells and CD19+ cells. Average mononuclear cell recovery following purging was 71% of the concentrated marrow with 78% yield of CFU-GM. Normal progenitor recovery related to patients' weight is predicted to be adequate for haemopoietic reconstitution following ablative chemoradiotherapy. This system is therefore feasible for large-scale clinical purging.


von Willebrand factor as a marker of endothelial cell activation following BMT. Endothelial cell activation may play a role in thrombotic complications of BMT such as hepatic veno-occlusive disease (VOD), right atrial line thrombosis and microangiopathic haemolysis. To assess this, von Willebrand factor antigen (vWF:ag) was measured in 72 patients (25 allografts, 46 autografts and one syngeneic) during the first 6 weeks post-transplant. There was a significant rise in vWF:ag in both allografts and autografts but a greater increase was seen in the allografts. The changes in vWF:ag did not correlate with changes in C reactive protein showing that this was not merely an acute phase response. vWF multimers were normal in a subgroup of uncomplicated transplants showing that there was no large scale endothelial cell disruption. Patients with VOD did not have changes in vWF:ag that were consistently different from uncomplicated controls. Three of four patients who developed line thrombosis had higher levels of vWF:ag compared with control groups; multimeric structure of the vWF was again normal. These results show that there is endothelial cell activation post-BMT and that this is greater in allografts compared with autografts, thus suggesting a possible mechanism for the higher incidence of VOD in this group. There were no useful predictive markers of VOD or thrombosis in individual patients.


Rapid and automated processing of bone marrow grafts without Ficoll density gradient for transplantation of cryopreserved autologous or ABO-incompatible allogeneic bone marrow. The growing number of BMTs has increased interest in safe and standardized in vitro bone marrow processing techniques. We describe our experience with a rapid automated method for the isolation of mononuclear cells (MNC) from large volumes of bone marrow using a Fenwal CS-3000 cell separator without employing density gradient materials. Forty bone marrow harvests with a mean volume of 1650 +/- 307 ml were processed. A mean of 75 +/- 34% (50 percentile range 54-94%) of the original MNCs were recovered in a volume of 200 ml with only 4 +/- 2% of the starting red blood cells (RBC). Removal of granulocytes, immature myeloid precursors and platelets proved to be sufficient to permit safe cryopreservation and successful autologous BMT (n = 25). Allogeneic BMT (n = 14, including three major ABO-incompatible) could be performed without additional manipulation. In both groups of patients timely and stable engraftment comparable to historical controls receiving Ficoll gradient processed autologous (n = 17) or unprocessed allogeneic BMT (n = 54) was observed. Moreover, 70 +/- 14% of the RBC could be recovered from the grafts. They were used for autologous RBC support of donors, rendering unnecessary autologous blood pre-donations.


Regimen-related toxicity in patients undergoing BMT with total body irradiation using a sweeping beam technique. In our institution, total body irradiation (TBI) is performed by means of a sweeping beam technique. Toxicity of the procedure was evaluated according to the only grading system designed for high dose chemoradiotherapy. One hundred patients undergoing TBI and conditioned with a standard cyclophosphamide regimen before BMT were evaluated. Regimen-related toxicity was graded according to the Seattle transplantation toxicity system, from 0 to IV (fatal toxicity), in eight organs on days 0, 7, 14, 28 and 100 for lungs. Eighteen patients did not develop any toxicity. Grades III, IV toxicities were uncommon (9%) and were not influenced by dose of TBI, GVHD prophylaxis, disease status and allogenicity although no grade IV toxicity was observed among autologous marrow recipients. However, grade II toxicity was more common in patients receiving allogeneic vs autologous grafts (p < 0.01) because of increased mucosal (p = 0.002) and liver (p = 0.12) toxicities. Renal toxicity was unevaluable. When cumulative toxicity was equal or higher than 4, day 100 survival was worse (p = 0.05). These data confirm the safety of our TBI procedure and the validity of the grading system except for renal toxicity. We suggest that a more aggressive conditioning regimen may be tolerated by patients receiving autologous grafts.


Reconstitution of the T cell receptor alpha beta repertoire in recipients of allogeneic BMT. This report describes the qualitative analysis of T cell receptor (TCR) repertoire regeneration in recipients of BMT. RNA samples from patient and control peripheral blood lymphocytes were prepared and tested for the presence of multiple V alpha and V beta transcripts by the polymerase chain reaction. TCR V gene expression was highly diverse within the first 6 months post-transplantation in recipients receiving either T cell-depleted or T cell-replete marrow, and in HLA mismatched as well as matched donor-recipient pairs. The sequencing of TCR message from BMT recipients also demonstrated J gene diversity and apparently normal junctional diversity at the V-J alpha join. Thus, T cell pools in BMT recipients are largely heterogeneous, not mono- or oligoclonal.


Erythropoietin treatment in allogeneic BMT accelerates erythroid reconstitution: results of a prospective controlled randomized trial. Twenty-eight allogeneic BMT patients (16 with acute leukemia, 12 with chronic myeloid leukemia) were included in a single center, prospective, randomized, controlled trial to assess the value of recombinant human erythropoietin (rh-Epo) in this setting. rh-Epo was administered through a central venous catheter as a single bolus injection (days 0-7: 100 U/kg/d; days 7-30: 150 U/kg/d). No secondary effects to rh-Epo treatment were detected. An earlier appearance of reticulocytes and a diminished need of red blood cells (RBCs) transfusions were observed in patients who were treated with rh-Epo (4 units vs 12 units; p < 0.05). The time to unsupported platelets above 25 x 10(9)/l was less in patients treated with rh-Epo than in control patients (19 days vs 31; p < 0.05), and they received significantly fewer platelet transfusions (36 units vs 138.5; p < 0.05). Our results show that rh-Epo treatment is capable of accelerating the erythroid reconstitution and decreasing the need for RBC transfusions. A beneficial effect on platelet reconstitution is also suggested, but further studies are necessary to confirm this point.


Acute suppurative thyroiditis complicating second allogeneic transplant for juvenile CMML. We describe here a 3-year-old boy with juvenile chronic myelomonocytic leukemia and monosomy 7 who underwent a second HLA matched bone marrow transplant from his sister. He developed hypothyroidism due to acute suppurative thyroiditis in the recovery phase of BMT. This is an extremely rare complication of BMT.


New method of limb deformities correction in children. A new "bloodless" technique (Ilizarov) was used to correct 36 limb deformities in 29 children. There were six leg length discrepancies, five achondroplasias, four deformed feet, five joint contractures, one rotational deformity of tibia, and in three the apparatus was used as an external fixator after corrective osteotomy. Lengthening was accomplished in 15 of the 16 procedures (93%). Average increase in femur length was 10 cm (32%), in tibial length 7.5 cm (30%), in humerus 11 cm (40%). Bony union was achieved in two out of five pseudoarthroses. Four deformed feet were fully corrected. Joint contractures were corrected in four out of five. The complication rate is as high as in other methods but with the Ilizarov apparatus, longer segments of bone were lengthened and more complex deformities were treated. Complications lessened as experience was gained.


Rectus sheath haematoma in a canoeist.

Rectus sheath haematoma in a canoeist. A 26-year-old male canoeist was referred with a 10-day history of abdominal pain, and a palpable mass in the left upper quadrant. No history of direct trauma was given. He was not taking any medication, and malignancy and inflammatory conditions were considered in the differential diagnosis. Ultrasonographic scan identified a mass originating in the rectus abdominis sheath. Ultrasonographically guided aspiration yielded some partially clotted blood, confirming the clinical diagnosis of rectus sheath haematoma. After conservative treatment, the patient resumed training, and is fully asymptomatic 1 year after discharge.


Understanding of Weil's disease among canoeists. Although the risks of contracting Weil's disease in the UK are small, it can be a serious illness. Recently the disease has attracted public interest. This interest has prompted confusion and anxiety among recreational water user groups such as canoeists who had not previously seen themselves as being particularly at risk. In this paper, the experience of symptoms associated with Weil's disease among canoeists, their understanding and perceptions of the disease, and their compliance with preventive advice on the British Canoe Union (BCU) warning card, have been studied. Gaps in understanding and of adherence to preventive advice were identified. Accordingly, it was concluded that health education for canoeists could be improved. Consideration is being given by the BCU to the value of preparing a canoeist instructors' training pack.


Daily physical activity and blood lactate indices of aerobic fitness in children. This study examined the relationship between daily physical activity and aerobic fitness in 11-16-year-olds. Habitual physical activity was assessed in 28 boys (mean(s.d.) age 13.6(1.3) years) and 45 girls (mean(s.d) age 13.7(1.3) years) from minute-by-minute heart rate monitoring during 3 school days. Aerobic fitness was assessed by determining the percentage peak VO2 at blood lactate reference values of 2.5 and 4.0 mmol l-1 during incremental treadmill running. The 4.0 mmol l-1 level occurred at a mean(s.d.) value of 89(7)% peak VO2 in both boys and girls and mean(s.d.) values at the 2.5 mmol l-1 level were 82(9)% peak VO2 in girls. Mean(s.d.) percentage time with heart rates at or above 140 beats min-1 was 6(3)% in boys and 5(3)% in girls. Corresponding values for percentage time at or above 160 beats min-1 were 3(2) for boys and 2(1) for girls. The number of 10- and 20-min periods of activity with the heart rate sustained above the 140 and 160 beats min-1 thresholds were also totalled over the 3 days. No significant relationships were identified between percentage peak VO2 at the 2.5 or 4.0 mmol l-1 blood lactate reference levels and either percentage time or number of 10- or 20-min periods above 140 or 160 beats min-1 (P > 0.05). These results support the hypothesis that daily physical activity levels in 11-16-year-old children do not stress aerobic metabolism sufficiently to influence aerobic fitness.


Training-overtraining: performance, and hormone levels, after a defined increase in training volume versus intensity in experienced middle- and long-distance runners. Performance and hormones were determined in eight middle- and nine long-distance runners after an increase in training volume (ITV, February 1989) or intensity (ITI, February 1990). Seven runners participated in both studies. The objective was to cause an overtraining syndrome. The mean training volume of 85.9 km week-1 increased within 3 weeks to 176.6 km week-1 during ITV and 96-98% of training volume was performed as long-distance runs at mean(s.d.) 67(8)% of maximum capacity. Speed endurance, high-speed and interval runs averaging 9 km week-1 increased within 3 weeks to 22.7 km during ITI, and the total volume increased from 61.6 to 84.7 km. A plateau in endurance performance and decrease in maximum performance occurred during ITV, probably due to overtraining, with performance incompetence over months. Nocturnal catecholamine excretion decreased markedly (47-53%), contrary to exercise-related plasma catecholamine responses, which increased. Resting and exercise-related cortisol and aldosterone levels decreased. Improvement in endurance and maximum performance occurred during ITI indicating a failure to cause an overtraining syndrome in ITI. Decrease in noctural catecholamine excretion was clearly lower (9-26%), exercise-related catecholamine responses showed a significant decrease, cortisol and aldosterone levels remained almost constant, exercise-related prolactin levels decreased slightly. There were no differences in insulin, C-peptide, free testosterone, somatotropic hormone (STH), follicle stimulating hormone (FSH), luteinizing hormone (LH), thyroid stimulating hormone (TSH), tri-iodothyronine (T3) and thyroxine (T4). The decrease in nocturnal catecholamine excretion during ITV might indicate a decrease in intrinsic sympathetic activity in exhausted sportsmen. But it remains open whether this reflected a central nervous system incompetence.


Retrospective survey of rugby injuries in the Leinster province of Ireland 1987-1989. Rugby Union Football is a high-speed contact sport and has a significant incidence of injury. This study evaluated senior rugby players with reference to the anatomical site of injury for each playing position.


Fracture of the lateral process of the talus: computed tomographic scan diagnosis. Fracture of the lateral process of the talus is rare but can be mistaken for a simple ankle sprain. A case with normal conventional radiographs is presented to draw attention to this diagnosis in the resistant ankle sprain, and to highlight some of the problems that may be encountered with treatment.


Acromioclavicular disruption in first class rugby players. In a random sample of 105 first class rugby players, 45% gave a history of injury of the acromioclavicular joint. All continued to play at the highest level. The effects of the injury appeared to be minimal. Supraspinatus impingement syndrome commonly associated with acromioclavicular pathology was sought, but not found.


A modified repair for the anterior cruciate ligament deficient knee. A retrospective analysis of 48 sportsmen and women from an original series of 76 consecutive patients who had undergone a modified McIntosh repair was carried out to establish whether or not the procedure could provide a satisfactory recovery and return to previous ability. The type and level of sport before injury was compared with that after operation. Symptoms of pain and giving way, and examination findings of pivot shift, and Lachman's test were compared before and after operation. Of the 48 patients assessed, 28 (58%) returned to full sporting capacity; 17 (35%) patients were participating in different sports or lower levels of their previous sports, and three patients did not participate in any sport. The more severely symptomatic knees did not perform so well after operation. The degree of preoperative anterior draw and Lachman's test did not influence the final result and the pivot shift, present in all before operation, was abolished in all but one case, which remained badly symptomatic. Concurrent meniscal injury or medial or lateral laxity did not influence return to sport once a full postoperative recovery was made. No deterioration was noticed in the level of sporting ability achieved thereafter. In this study it has been shown that the modified McIntosh repair is a swift extra-articular reconstruction for the anterior cruciate ligament deficient knee, which is less elaborate than previously described Ellison and McIntosh procedures, and which has produced a comparable result.


Injuries of the spine sustained during rugby. A comparison was made between the number of rugby injuries seen between 1956 and 1982 (67) and the number of rugby injuries seen between 1982 and 1987 (20). The standards, the positions, the mechanics of injury and the fitness of the players were analysed. It was concluded that the law changes had resulted in a dramatic fall in the number of players injured, that it was the less fit and less skilled players that were getting injured, and that the laws were adequate but were not being enforced.


Dying to be big: a review of anabolic steroid use. Anabolic steroids use is commonly perceived to be the domain of the higher echelons of competitive athletes. However, a great deal of anabolic steroid use occurs in private gymnasia (non-local authority) among non-competitive recreational athletes. Our study has attempted to give an insight into the prevalence of the use of these drugs, the hazards associated with it, and the public health responses which we have adopted.


Physiological profiles of Hong Kong élite soccer players. Most physiological profiles of élite soccer players originate from Western Europe and North America. Unfortunately, there is a scarcity of descriptive data on the physical characteristics of Asian soccer players. Therefore, the purpose of this study was to evaluate the physiological profiles of élite soccer players in Hong Kong. It was conducted in conjunction with the selection of the Hong Kong team before the 1990 Beijing Asian Games. In all, 24 professional soccer players were selected from a pool of 180 players as subjects for the study. The following means(s.d.) were observed: height 173.4(4.6) cm; weight 67.7(5.0) kg; body fat 7.3(3.0)%; forced vital capacity (FVC) 5.1(0.6) l; maximum oxygen uptake (VO2max) 59.1(4.9) ml kg-1 min-1; anaerobic threshold (AT 80.0(7.2)% of VO2max; alactic power index 13.5(2.4) W kg-1; lactic work index 298(27) J kg-1; peak isokinetic dominant knee extensor and flexor strengths 2.72(0.36) Nm kg-1 and 1.65(0.20) Nm kg-1. On average the physique of Hong Kong soccer players appeared to be smaller and lighter than those found in Europe, which may be one of the key factors that contribute to the lack of success of Hong Kong soccer teams in international competition.


Muscle soreness, swelling, stiffness and strength loss after intense eccentric exercise. High-intensity eccentric contractions induce performance decrements and delayed onset muscle soreness. The purpose of this investigation was to study the magnitude and time course of such decrements and their interrelationships in 26 young women of mean(s.d.) age 21.4(3.3) years. Subjects performed 70 maximal eccentric contractions of the elbow flexors on a pulley system, specially designed for the study. The non-exercised arm acted as the control. Measures of soreness, tenderness, swelling (SW), relaxed elbow joint angle (RANG) and isometric strength (STR) were taken before exercise, immediately after exercise (AE), analysis of variance and at 24-h intervals for 11 days. There were significant (P < 0.01, analysis of variance) changes in all factors. Peak effects were observed between 24 and 96 h AE. With the exception of STR, which remained lower (P < 0.01), all variables returned to baseline levels by day 11. A non-significant correlation between pain and STR indicated that pain was not a major factor in strength loss. Also, although no pain was evident, RANG was decreased immediately AE. There was no relationship between SW, RANG and pain. The prolonged nature of these symptoms indicates that repair to damaged soft tissue is a slow process. Strength loss is considered particularly important as it continues when protective pain and tenderness have disappeared. This has implications for the therapeutic management of patients with myopathologies and those receiving eccentric exercise for rehabilitation.


Spondylolysis in fast bowlers: principles of prevention and a survey of awareness among cricket coaches. Back injury in general, and spondylolysis in particular, represents a serious threat to the fast bowler. Hereditary factors, poor technique, overuse, and poor preparation for fast bowling may combine to produce the 'at risk' bowler. Three first-class county coaches were interviewed to establish a level of awareness of the lesion, with an emphasis on preparation of fast bowlers and the roles of technique alteration and bowling prescription in reducing the risk of back injury.


Effect of caffeine on maximal strength and power in élite male athletes. Computerized testing of 20 élite male athletes was performed to determine the effect of 7 mg kg-1 caffeine on strength and power of the knee extensors and flexors. Subjects received counterbalanced administrations of either caffeine or a placebo on two separate occasions. Peak torque (T) was measured for knee extension (ET) and flexion (FT) at angular velocities of 30 degrees, 150 degrees and 300 degrees s-1. Additionally, performance for the first 125 ms (TAE) and power (W) were recorded during 300 degrees s-1. Testing sessions were held 1 week apart, at which time the placebo/caffeine administration was reversed. A 2 x 2 repeated measures analysis of variance supplemented with a Neuman-Keuls post hoc test showed the following--significant caffeine-related increases (P < 0.05) for ET at 30 degrees s-1, ET at 300 degrees s-1, and ETAE, and EW at 300 degrees s-1. Dependent t-tests performed for pre- to post-test means showed significant changes for the caffeine group in ET at 30 degrees s-1, FT at 30 degrees s-1, FT at 150 degrees s-1, ET at 300 degrees s-1, FT at 300 degrees s-1, E and FTAE, and EW at 300 degrees s-1. No significant effects were found for the placebo trial in any variable. It was concluded that caffeine can favourably affect some strength parameters in highly resistance-trained males. However, differences in subject fibre type, motivation and caffeine sensitivity need to be elucidated.


[Auto-adaptative network separator of sources]. This article defines a new convergence criterion for the modelization of networks with modifiable synapses, as presented by Jutten, Herault, Ans ([1], [2]). The network adaptation law, empirically determined in [1], [2], is reconsidered: a theoretical law concerning the evolution of system coefficients and outputs is demonstrated and validated on simulations. The system always converges towards a unique attractor, as predicted by the theoretical convergence criterion. This system evolution towards a particular stable state enhances the interesting physical aspect of this modelization.


[Morphologic differentiation of modern humans: correspondence between skull form and geographic distribution of population]. Multivariate distance analysis of 7 to 9 classical craniometric dimensions was carried out on a large number (n = 536) of modern human populations. Results obtained showed that the scatter plot based on morphological affinities was very similar to geographic dispersion of sample populations. Asia appeared to be closer to a possible center of dispersion than any other location.


Materials management considerations in critical care areas. The materials management department has the potential to provide great value to the nursing department in the planning, relocation, and occupation of new patient care areas. Functional responsibilities in the materials area compels these personnel to evaluate issues such as contractor compliance; new products and equipment on the market; price trends; required lead times; traffic patterns inside the hospital; space requirements for clean, soiled, and hazardous materials; utilization patterns of supplies and equipment; and shifting demands on the time of the clinical staff.


Brain-stem anesthesia as a complication of regional anesthesia for ophthalmic surgery. Brain-stem anesthesia is a serious complication of orbital regional anesthesia that may occur when the injected local anesthetic agent gains access to the central nervous system by direct spread from the apex of the orbit via submeningeal pathways. In most studies the reported incidence rate during retrobulbar block is one case per 350 to 500 patients. Failure to recognize the condition or to treat it adequately may be life threatening. Treatment includes reassurance, intravenous administration of fluids, pharmacologic circulatory support or suppression of convulsions, and cardiopulmonary resuscitation. With proper treatment complete recovery is the rule. In all situations in which orbital block is to be done, basic cardiopulmonary resuscitation equipment and personnel familiar with its use are essential. Monitoring of the blood pressure, electrocardiography and pulse oximetry should be routine. Having the globe in primary gaze renders the optic nerve less vulnerable, and avoidance of deep penetration of the orbit is of great importance.


Trans-scleral neodymium: YAG retinal photocoagulation in rabbit eyes. To assess the ability of trans-scleral application of the continuous-wave neodymium: YAG laser to produce retinal ablation with reproducible chorioretinal scars, retinal photocoagulation with the YAG laser was done in four Dutch-belted rabbits (eight eyes). Sharply defined, reproducible lesions were produced that were identical to those found following conventional treatment with the argon laser. At 0.2 J there was destruction of the outer retinal layers with sparing of the inner retinal layers. At 3.0 J full-thickness retinal reorganization was noted; however, even at this energy setting there was minimal histologic evidence of scleral damage. Focused laser energy produced distinctly more retinal destruction and obliteration of the retinal pigment epithelium than unfocused energy. The results suggest that trans-scleral photocoagulation with the YAG laser may be possible and would be particularly applicable in situations in which media opacities preclude visualization of the retina.


Cell culture of macular epiretinal membranes. To attempt to differentiate the cells of origin in epiretinal membranes, cells were cultured from an idiopathic epiretinal membrane and an epiretinal membrane that formed after successful repair of a retinal detachment. The cells exhibited a variety of forms. When the cells from the idiopathic epiretinal membrane were incubated with melanin from donor eyes a population of small cells accumulated the pigment, but the large, flat cells did not. The epiretinal membrane was not composed of a single cell type. Although a population of cells exists within epiretinal membranes, no one cell type could be identified as the cell type of origin.


Stability of fixation in healthy subjects during automated perimetry. The stability of fixation of 12 healthy young subjects was assessed with a high-resolution eye tracker during automated perimetry. All the subjects exhibited eye movements and fixation shifts during the examination. The average shift in eye position between successive 1-minute intervals ranged from 0.41 degrees to 2.11 degrees, and the average standard deviation of eye position around the average of each interval ranged from 0.56 degrees to 2.55 degrees. In nine of the subjects the eye remained within 3 degrees of centre for 96% or more of the stimuli. The remaining three subjects experienced difficulty maintaining central fixation: the eye was more than 3 degrees away from centre for 28% or more of the stimuli. In the latter subjects the ability to repeatedly stimulate the intended retinal test locations was substantially reduced. The authors discuss the effects of unstable fixation on short-term fluctuation and on fixation loss.


An ocular dynamic study supporting the hypothesis that hypothyroidism is a treatable cause of secondary open-angle glaucoma. To examine the hypothesis that glaucoma may be a manifestation of unrecognized hypothyroidism, we studied 25 consecutive patients with newly diagnosed hypothyroidism who presented to an endocrinology clinic. Using tonography and tonometry, we demonstrated a reduction in facility of outflow in the hypothyroid state. With treatment of the hypothyroidism alone there was a statistically significant improvement in facility of outflow, intraocular pressure and Po/C (p < 0.002). Our results support the hypothesis that secondary open-angle glaucoma may be a manifestation of hypothyroidism and that the glaucoma will resolve on treatment of the primary disease.


Reversal of poorly controlled glaucoma on diagnosis and treatment of hypothyroidism. Hypothyroidism was diagnosed in a 62-year-old woman with poorly controlled primary open-angle glaucoma, including a disc hemorrhage and a documented arcuate scotoma. After 1 year of thyroxine therapy the glaucoma was easily controlled, pilocarpine treatment was stopped and the visual fields were completely normal. To our knowledge this is the first report in the modern literature of reversal of open-angle glaucoma with treatment of hypothyroidism.


Favourable outcome of traumatic endophthalmitis with associated retinal breaks or detachment. Traumatic endophthalmitis in association with retinal breaks or detachments is reported to have uniformly poor visual and anatomic outcomes. We describe two cases of culture-positive traumatic endophthalmitis with retinal breaks or detachment in which the final visual result was 20/70 in one case and 20/40 in the other. Factors that may improve the prognosis in such cases include attention to the possibility of infection, selective use of broad-spectrum antibiotics, prompt surgical intervention and improvement in vitreous surgical techniques. Modification of intravitreal antibiotic regimens may be indicated in eyes in which the vitreous cavity is partially filled with air or gas.


Periorbital hyperpigmentation and erythema dyschromicum perstans. Erythema dyschromicum perstans is a rare idiopathic dermatosis characterized by ash-grey, well-demarcated skin lesions, which may involve the face. We describe an 8-year-old girl with erythema dyschromicum perstans presenting as bilateral acquired periorbital hyperpigmentation. The changes seen on histologic study of a skin biopsy specimen were consistent with the clinical diagnosis. The various causes of periorbital hyperpigmentation and characteristics of erythema dyschromicum perstans are reviewed.


Role of sodium and water excretion in the antihypertensive effect of vasopressin in the spontaneously hypertensive rat. Mean arterial pressure (mmHg (1 mmHg = 133.322 Pa)), sodium excretion rate (mumol.kg-1.min-1), and urine flow (microL.kg-1.min-1) were measured in conscious unrestrained spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto rats (WKY) before, during, and after a 3-h intravenous infusion of arginine vasopressin (20 ng.kg-1.min-1), an equipressor dose of phenylephrine, or an infusion of the vehicle. Cessation of the phenylephrine infusion was associated with a return of arterial pressure to preinfusion control values in both SHR and WKY. Cessation of the vasopressin infusion was also associated with a return of arterial pressure to preinfusion values in WKY. In contrast, in the SHR, arterial pressure fell from a preinfusion control level of 164 +/- 6.2 to 137 +/- 4 mmHg within 1 h of stopping the vasopressin infusion. Five hours after stopping the infusion, pressure was 134 +/- 3 mmHg (29 +/- 5 mmHg below preinfusion levels). Similar to the WKY, cessation of a vasopressin infusion was associated with a return of arterial pressure to preinfusion values in Sprague-Dawley rats. Thus, the failure to observe a hypotensive response in normotensive rats was not a peculiarity of the WKY strain. Sodium excretion rates increased during the infusions of vasopressin to a greater extent in SHR than in WKY. However, the natriuresis induced by phenylephrine was not significantly different from that generated by vasopressin in SHR, and in WKY, the natriuresis was greater for phenylephrine than for vasopressin. Urine output increased to a greater extent during the infusions of phenylephrine in both SHR and WKY than during vasopressin infusion.(ABSTRACT TRUNCATED AT 250 WORDS)


Differences in myocardial ischemic tolerance between 1- and 7-day-old rabbits. Between 1 and 7 days of life, the newborn rabbit heart shifts from predominantly using carbohydrates to predominantly using fatty acids as an energy substrate. We therefore used isolated working hearts from 1- or 7-day-old rabbits to determine the effects of fatty acids on myocardial glucose use and the ability of hearts to recover following various periods of transient no-flow ischemia. One-day-old hearts were perfused via the inferior vena cava and ejected buffer through the cannulated aorta and pulmonary artery. Seven-day-old hearts were perfused via the left atrium and ejected buffer through the cannulated aorta. To measure glucose use, hearts were perfused with 11 mM [3H, 14C]glucose, 3% albumin, and 500 microU insulin/mL, in the presence or absence of 0.4 mM palmitate. In the absence of fatty acids, glycolytic rates were similar in 1- and 7-day-old hearts, whereas glucose oxidation rates were 5 times greater in 7-day-old hearts. Palmitate did not have any major effects on overall glucose use in 1-day-old hearts, but did markedly inhibit glycolysis and glucose oxidation in 7-day-old hearts. A series of hearts were also subjected to periods (25-60 min) of no-flow ischemia, followed by 30 min of aerobic reperfusion. In the absence of palmitate, 1-day-old hearts subjected to ischemic periods of up to 60 min recovered some degree of mechanical function during reperfusion, whereas 7-day-old rabbit hearts failed to recover if hearts were subjected to ischemic periods of 35 min or longer.(ABSTRACT TRUNCATED AT 250 WORDS)


Recovery of rat tibialis anterior motor unit properties following partial denervation. The recovery of selected mechanical, morphological, and metabolic properties of rat tibialis anterior fast motor units was determined following partial denervation (n = 7) or partial denervation and hemispinal cord transection (n = 5) and compared with age-matched control units (n = 7). Following 1-12 months of recovery, the mechanical properties of each unit were measured and the fibres depleted of glycogen by using standard ventral root filament stimulation techniques. Quantitative histochemical techniques were used to determine cross-sectional area and the activities of succinate dehydrogenase and alpha-glycerolphosphate dehydrogenase in individual unit fibres. Partial denervation increased the mean fibre area but decreased alpha-glycerolphosphate dehydrogenase activity. Succinate dehydrogenase was unchanged in the denervated groups. The variability in area and enzymatic activities among the unit fibres was unchanged. However, the interrelationship between the enzymes was altered by both denervation procedures. Succinate dehydrogenase activity was directly related to fatigue resistance and inversely related to tetanic tension across the units. These findings suggest that a motor unit reestablishes many of its properties despite marked changes to the composition of the unit brought about by partial denervation. In addition, a reduction in the neuromuscular activity of units during reorganization had a limited effect on recovery.


Antioxidant changes in heart hypertrophy: significance during hypoxia-reoxygenation injury. Because hypertrophied rat hearts display an increase in antioxidant enzyme activities and because hypoxia-reoxygenation injury is known to involve free radicals, we tested the hypothesis that the hypertrophied heart may be more resistant to this type of injury. Hypertrophied rat hearts after 10 weeks of chronic pressure overload showed elevated superoxide dismutase (SOD) and glutathione peroxidase (GSHPx) activities and a decrease in lipid peroxidation as indicated by malondialdehyde (MDA) content. Glucose-free hypoxia for 15 min resulted in a complete failure of developed tension and about 200% increase in resting tension in both hypertrophied and sham control groups (p < 0.05). Upon reoxygenation for up to 30 min, hypertrophied hearts recovered developed tension to 60% and resting tension was higher by only 80% of prehypoxic values. In contrast, sham hearts showed only a 25% recovery of developed tension, whereas resting tension remained 130% higher than prehypoxic control values. During hypoxia, the SOD activity was significantly reduced in both sham and hypertrophied groups, whereas GSHPx was reduced only in the sham group. Upon reoxygenation there was no further change in these enzyme activities. Both the SOD and GSHPx activities in the hypertrophied group remained significantly higher than the corresponding reoxygenated sham hearts. During hypoxia, there was no apparent change in MDA content in either the sham or hypertrophied hearts. However, reoxygenation resulted in a significant increase in MDA content in both sham and hypertrophied hearts, but the MDA content was significantly less in the hypertrophied group (p < 0.05). It is suggested that maintenance of an adequate endogenous antioxidant reserve during hypoxia may be important in recovery upon reoxygenation.


Role of environmental temperature and photoperiod in regulation of seasonal testicular activity in the frog, Rana perezi. To analyze the role of environmental temperature and photoperiod in the regulation of the annual testicular cycle in Rana perezi, we performed experiments combining high (25 +/- 1 degrees C) or low (6 +/- 1 degrees C) temperature and different photoperiod regimens (18L:6D, 12L:12D, and 6L:18D (hours light:hours dark)) during three phases of the reproductive cycle: winter stage (December) and prebreeding (February) and postbreeding (May, June) periods. Low temperature and short photoperiod in winter induced the arrest of the maturation phase of spermatogenesis and the activation of primary spermatogonia proliferation and spermiohistogenesis. Rana perezi testis responded to long days stimulus in winter, even at low temperature, with induction of the maturation phase of the cycle. Exposure of male frogs to either high temperature or long photoperiod induced a decrease in testosterone levels in winter. During the prebreeding period, an increase in environmental temperature caused a reduction in testosterone, and a lengthening in photoperiod produced the opposite effect. Photoperiod had no effect on testosterone levels during the postbreeding period, but low temperature increased testosterone plasma levels. These results suggest that both temperature and photoperiod effects can vary seasonally, depending on the phase of the annual reproductive cycle in R. perezi.


Muscle metabolism in track athletes, using 31P magnetic resonance spectroscopy. We tested whether preferred running event in track athletes would correlate with the initial rate of phosphocreatine (PCr) resynthesis following submaximal exercise. PCr recovery was measured in the calf muscles of 16 male track athletes and 7 male control subjects following 5 min of repeated plantar flexion against resistance. Pi, PCr, and pH were measured using phosphorus magnetic resonance spectroscopy (31P MRS) with an 8-cm surface coil in a 1.8-T magnet. During exercise, work levels were gradually increased to deplete PCr to 50-60% of the initial value. No drop in pH was seen in any of the subjects during this exercise. The areas of the PCr peaks following exercise were fit to monoexponential curves. Two or three tests were performed on each subject and the results averaged. Athletes were divided into three groups based on their primary event: sprinters running 400 m or less, middle-distance athletes running 400-1500 m, and long-distance athletes running farther than 1500 m. The maximal rates of PCr resynthesis (mmol.min-1.kg-1 muscle weight) were 64.8 +/- 8.6, for long-distance runners; 41.4 +/- 11, for middle-distance runners; 32.0 +/- 7.0, for sprinters; and 38.6 +/- 10, for controls (mean +/- SE). The faster PCr recovery rates seen in long-distance runners compared with sprinters indicate greater oxidative capacity, which is consistent with the known differences between athletes in these events.


Acute administration of bromocriptine abolishes the hyperprolactinemic response induced by submaximal exercise in man. The effective control of hypophysial prolactin (PRL) secretion with a pharmacological agent is a prerequisite for the investigation of the role of hyperprolactinemia observed during exercise. Using bromocriptine, a potent inhibitor of PRL secretion, this study established the proper experimental conditions whereby any significant increase in plasma PRL level can be prevented and basal circulating levels maintained during physical exercise. On three occasions at weekly intervals, 15 male adults, separated into two groups, exercised on an ergocycle (40 min at 65% VO2max) either 1 or 3 h after ingesting either placebo or 1.25 or 2.50 mg of bromocriptine mesylate (Parlodel; Sandoz Canada Inc., Dorval, Qué.). Under all conditions, the plasma PRL elevation observed during exercise after placebo was prevented by the administration of bromocriptine. Resting plasma PRL levels were maintained when exercise was performed 1 h after bromocriptine ingestion, but were significantly reduced when exercise was performed 3 h after administration of either bromocriptine dosages. Considering the primary and secondary effects observed, 1.25 mg of bromocriptine administered 1 h before exercise provides suitable experimental conditions to investigate the role of the increase in plasma PRL during physical exercise.


Moderate increases in peripheral blood estradiol concentration in the adult ram do not directly inhibit testosterone secretion. Two experiments were conducted in July with adult Dorset x Leicester x Suffolk rams to determine whether increases of 150 or 300% in estradiol (E2) concentration in peripheral blood (from 6.3 +/- 0.8 pg/mL in control rams) would affect testosterone secretion directly as well as indirectly via the hypothalamic-pituitary axis. After 4 days of estradiol treatment (experiment 1) provided with subcutaneous polydimethylsiloxane implants filled with crystalline estradiol, luteinizing hormone (LH) and testosterone secretions were reduced by 50% (p < 0.05) in both groups of rams because of subtle decreases in pulse frequencies and amplitudes. Estradiol treatments were also associated with decreases in mean follicle-stimulating hormone (FSH) concentration (30-50% in both groups, p < 0.05) and increases in mean prolactin concentration (35% in low-E2 group; 105% in high-E2 group, p < 0.05), but testicular responsiveness to an LH challenge (single intravenous dose, 10 micrograms NIH-LH-S25) remained normal. When along with estradiol treatment, 10-micrograms doses LH were given every 80 min (experiment 2), testosterone secretion increased by 265% (p < 0.05) in both treated and control rams. Relative to day -1, secretion on day 4 was characterized by higher (p < 0.05) pulse frequencies and baseline concentrations and lower (p < 0.05) pulse amplitudes; values for all characteristics were similar to those for Dorset x Leicester x Suffolk rams in the breeding season. Interestingly, the decreases in mean FSH concentration brought about by estradiol and (or) LH treatments were not any greater than in experiment 1, and estradiol's ability to elevate mean prolactin concentration was blocked completely.(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of acute swimming exercise on muscle and erythrocyte malondialdehyde, serum myoglobin, and plasma ascorbic acid concentrations. Acute exercise may induce free-radical production in mitochondria during basal metabolism of aerobic cells. Ascorbic acid is a strong antioxidant agent, whereas myoglobin is known to act as an oxygen reservoir. The purpose of this study was to investigate the effects of acute exercise on malondialdehyde levels in gastrocnemius, vastus medialis, and triceps brachi muscles and erythrocytes. In addition, we investigated the ascorbic acid levels and serum myoglobin concentrations in rats, following acute swimming exercise. We found that the levels of muscle malondialdehyde and serum myoglobin increased and the levels of plasma ascorbic acid decreased, in proportion to the duration of exercise; however, the levels of erythrocyte malondialdehyde did not change.


Atrial natriuretic peptide inhibits compensatory responses when cardiac performance is depressed. We tested the hypothesis that the atrial natriuretic peptide (ANP) mediated decrease in baroreceptor sensitivity that is seen in normal rats is more pronounced in a state of depressed cardiac performance. Holtzman rats (n = 15) were injected with Adriamycin (1 mg/kg i.p. 3 times/week for 8-10 weeks). Control rats (n = 17) were injected with 0.9% saline. Experiments were done in conscious animals that had been catheterized for i.v. infusions and for measurement of arterial blood pressure (ABP) and heart rate (HR). ANP (250 ng.kg-1.min-1) or saline vehicle was infused i.v. Graded periodic bolus injections of phenylephrine or sodium nitroprusside were given to assess baroreceptor sensitivity (beats.min-1.mmHg-1) up to 60 mmHg (1 mmHg = 133.3 Pa) above and below resting ABP. The following day the experiment was repeated with the ANP-vehicle regimen reversed. Finally, the rats were anesthetized and the rate of left ventricular pressure increase (dP/dt) was measured. Data evaluation included calculation of least squares linear regression slopes of peak delta HR vs. peak delta ABP, applying corrections for experimental errors in both the dependent and independent variables. Adriamycin rats (A) did not differ significantly from control rats (C) with respect to either initial ABP (A = 105 +/- 5; C = 100 +/- 3; mean mmHg +/- SEM) or initial HR (335 +/- 9 vs. 312 +/- 13 beats.min-1). However, their indices of cardiac performance were significantly depressed.(ABSTRACT TRUNCATED AT 250 WORDS)


Inhibitory and excitatory mechanisms of neurotensin action in canine intestinal circular muscle in vitro. The effect of neurotensin on canine ileal circular muscle devoid of myenteric plexus was investigated using single and double sucrose gap techniques. Similar results were obtained with microelectrode techniques. Neurotensin caused a temperature-sensitive and dose-dependent biphasic response, an initial hyperpolarization associated with inhibition of contractile activity, followed by an excitatory phase, usually consisting of spike discharge and tonic and phasic contractions, for which depolarization was not required. Neither response was affected by tetrodotoxin, phentolamine, propranolol, or atropine. The hyperpolarization was associated with decreased membrane resistance, blocked by 10(-7) M apamin, and converted to tonic depolarization by apamin (10(-6) M). Tachyphylaxis to neurotensin occurred when the stimulation interval was less than 20 min. After Ca2+ depletion, depolarization was observed instead of the hyperpolarization; this depolarization was not affected by nitrendipine and was gradually abolished with repetitive stimulation at 20-min intervals. When Ca2+ was present, nifedipine did not alter the hyperpolarizing phase of the response but inhibited spiking and blocked all contractions. The excitatory phase of the response was enhanced by Bay K-8644. Neuromedin N elicited a response identical with that of neurotensin. The responses of the two peptides were completely cross tachyphylactic. Inhibitory junction potentials were not affected by neurotensin tachyphylaxis. It is concluded that neurotensin and neuromedin N activate apamin-sensitive, calcium-dependent potassium channels in circular muscle, causing membrane hyperpolarization and inhibition of muscle contraction. Release of intracellular calcium is involved in the activation of these potassium channels.(ABSTRACT TRUNCATED AT 250 WORDS)


Portal venous infusions of L-glutamine in anaesthetized dogs do not influence renal function. It has been reported that the intraportal infusion of glutamine in Munich-Wistar rats will cause depression of renal perfusion and the urinary excretion of salt and water. We have attempted to reproduce these findings in anaesthetized dogs. L-Glutamine was infused at doses between 120 and 150 mumol/min into the portal vein and femoral vein of anaesthetized dogs. No effect was observed on portal venous pressure, blood pressure, or kidney function. Similar data were obtained with D-glutamine. Liver biopsy revealed no abnormalities. When 1.5-3 micrograms histamine (free base) was infused into the portal system, portal venous pressure rose from 15.2 +/- 0.33 to 24.8 +/- 0.40 cmH2O (p < 0.05) (1 cmH2O = 98.1 Pa). Glutamine infusions do not appear to initiate hepatorenal reflexes in dogs as they have been reported to do in rats.


[A career in nursing sciences?].

[A career in nursing sciences?]. A research study was conducted in a francophone high school in Northern Ontario to examine students' perceptions of nursing and the influence of these perceptions on nursing as a career choice. All students in grades 11, 12 and 13 were invited to participate. Fifty-eight percent (n = 268) completed the questionnaire. Results showed that 37 percent of the respondents considered pursuing a career in the health sciences. Only 14% percent were interested in nursing. Respondents' comments suggest that the nurse is viewed favorably but the profession is perceived as a career that does not involve pleasant tasks, good working conditions or opportunities for professional advancement. Reasons advocated for choosing nursing were altruistic rather than career-oriented. Students saw nursing practice as occurring mainly in a hospital setting. Half of the respondents who had chosen nursing as a career opted to enroll in a university program and the other half chose a college program. Results suggest that nursing continues to face an image problem regarding its role in the health care system. In these times of job losses and budget cuts, the profession still needs to attract young recruits. This is the challenge we have to face.


Preproenkephalin RNA increases in the hypothalamus of rats stressed by social deprivation. 1. Pharmacological evidence indicates that stress induced by brief (14 to 20-day) social deprivation in the rat is associated with an activation of the central preproenkephalin (ENK) opioid system. This study examines the neurochemical evidence that substantiates such an activation. 2. Using a specific ENK complementary DNA probe, ENK RNA levels were measured by dot blot and Northern blot analyses in different brain areas of socially deprived rats. Immunoreactivity to met-enkephalin-derived peptides was also evaluated by radioimmunoassay in the same brain regions. 3. Brief social deprivation increased the levels of ENK RNA and enkephalin immunoreactivity in whole hypothalamus. 4. Our data suggest that this type of stress appears to be associated to an induction of ENK gene transcription in hypothalamus.


Thyroidal influence on the cell surface GM1 of granule cells: its significance in cell migration during rat brain development. 1. No difference was observed in the in vitro growing ability of granule cells isolated from hypothyroid or normal rat brain. When granule cells were taken from hypothyroid rat brain and grown in normal culture medium containing 10% fetal calf serum, they behaved similarly to the granule cells obtained from normal rat brain. 2. In both cases there were progressive losses of in vitro growing ability of the granule cells with the age of the animal and it became impossible to grow them when derived from 21 days or older animals. 3. A marked decrease in cell surface GM1 was observed when the cells were maintained under thyroid hormone-deficient conditions in culture. 4. Anti-GM1 antibody was found to inhibit significantly the migration of granule cells along the astrocyte fibers. 5. These results indicate that GM1 has an important role in thyroid hormone-dependent postnatal brain maturation in rat.


Effects of chronic antidepressant treatment on dopamine-related [3H]SCH 23390 and [3H]spiperone binding in the rat striatum. 1. The effects of chronic administration of antidepressants on dopamine-related [3H]SCH 23390 and [3H]spiperone binding to rat striatal membranes were assessed. 2. The monoamine oxidase inhibitors phenelzine (5 or 10 mg kg-1/day) and tranylcypromine (1 mg kg-1/day) and the tricyclic desipramine (10 mg kg-1/day) were administered for 28 days by constant subcutaneous infusion using Alzet (2ML4) osmotic minipumps. 3. These treatments did not alter Kd estimates for either [3H]SCH 23390 or [3H]spiperone binding sites. The monoamine oxidase inhibitors induced a decrease in the Bmax values for both [3H]SCH 23990 and [3H]spiperone binding sites. Desipramine induced a decrease in the Bmax value for [3H]SCH 23390 binding but had no effect on the Bmax value for [3H]spiperone binding.


DNA polymerase alpha--primase complex of Physarum polycephalum. DNA polymerase alpha and DNA polymerase alpha--primase complex of Physarum polycephalum were purified by rapid methods, and antibodies were raised against the complex. In crude extracts, immune-reactive polypeptides of 220 kDa, 180 kDa, 150 kDa, 140 kDa, 110 kDa, 86 kDa, 57 kDa and 52 kDa were identified. The structural relationships between the 220 kDa, 110 kDa and 140 kDa (the most abundant form) was investigated by peptide mapping. The 140 kDa form was active DNA polymerase alpha. The 57 kDa and the 52 kDa polypeptides were identified as primase subunits by auto-catalytic labelling. In amoebae, the immune-reactive 140 kDa polypeptide was replaced by a 135 kDa active DNA polymerase alpha.


Effects of tubulozole on the amoeboflagellate transformation in Physarum polycephalum. The Amoeboflagellate Transformation (AFT) of Physarum polycephalum involves rapid changes in the cytoskeleton, cell shape and cell motility. Use of pharmacologic agents to probe the role of cytoskeletal elements in the AFT are impeded because the transforming cells are very sensitive to such commonly-used drug solvents as DMSO. The anti-microtubule agent tubulozole is found to disrupt, rapidly and transiently, the AFT, inhibiting flagella formation, cell elongation and the arrangement of microtubules and microfilaments. Cells recover quickly, possibly due to precipitation of the drug; the reappearance of normal arrays of microfilaments and cytoplasmic microtubules lags behind flagella formation.


Cellular and molecular analysis of plasmodium development in Physarum. The development of an amoeba into a plasmodium involves extensive changes in cellular organisation and gene expression. The genetic basis of a number of recessive mutations that block plasmodium development has been elucidated. The stage at which development becomes abnormal has been determined for all the mutants, as has the terminal phenotype. In order to investigate the changes in gene expression that accompany plasmodium development, a cDNA library has been made using RNA isolated from cell populations in which development was occurring.


[Protective effect of sho-saiko-to (TJ 9) in experimental liver injury]. Experiments carried out on male mice (ICR) demonstrated a protective effect of premedication with the preparation Sho-Saiko-To (TJ 9, Tsumura and Comp.) against the hepatotoxic effects of CCl4 and T1-acetate, which were manifested by increased peroxidation of lipids and increased depletion of reduced glutathion in liver homogenates.


[The effect of kampo preparations and their effect on peptidase activity in damage caused by free radicals]. The activity of peptidases (trypsin from bovine pancreas and trypsin-like enzymes from the liver rat homogenate) was influenced by five preparation of Kampo medicine, TJ-9 (Sho-Saiko-To), TJ-15 (Oren-Gedoku-To), TJ-23 (Toki-Shakuyaku-San), TJ-96 (Saiboku-To), and TJ-114 (Sairei-To) and studied in relation to their effect on the uptake of free oxygen radicals demonstrated earlier. On the basis of increased activity of trypsin and trypsin-like enzymes and the previously found capability of uptaking free oxygen radicals, the mechanism of action of the Kampo preparations may be assumed to be connected not only with a direct support of enzymes of digestion and increased activity of peptidases, capable of eliminating oxidatively damaged proteins, but with an antioxidative effect as well, which prevents increased cumulation of oxidatively damaged macromolecules and the action of superoxide radicals developed earlier by the well-known and trypsin-stimulated conversion of xanthinedehydrogenase to xanthinoxidase.


[Antioxidative activity of oren-gedoku-to (TJ-15) and toki- shakuyaku-san (TJ-23) in the liver microsomes of rats]. TJ-15 (Oren-Gedoku-To) inhibited enzymatically (NADPH or CumOOH) and non-enzymatically (Fe-askorbate) induced lipid peroxidation in the rat liver microsomes as assessed by the TBA-reactive product accumulation. TJ-23 (Toki-Shakuyaku-San) had little effect on either system. The protective effect of TJ-15 against lipid peroxidation could not be fully accounted for by its action on microsomal electron transfer, as evaluated by studying the kinetics of reduction of cytochrome C. In a free solution TJ-15 and TJ-23 effectively scavenged OH., radicals, as indicated by the inhibition of ethylene production from KMBA, and O2-. anion radicals, as assessed by the inhibitory effect on the rate of NBT reduction. The present results suggest that TJ-15, yet not TJ-23, is capable to reach hydrophobic intra-membrane sites at concentrations at which it is an effective antioxidant. Thus TJ-15 may be a potentially useful protective agent against free radical-mediated damage.


[Antioxidative properties of the kampo drugs TJ-9, TJ-15, TJ-23, TJ-96 and TJ-114]. Plant extracts from Kampo medicines TJ-9 (A), TJ-15 (B), TJ-23 (C), TJ-114 (D) and TJ-96 (E) inhibited peroxidation of low-density lipoprotein (LDL) in vitro and peroxidation of multilamellar liposomes formed by total lipids isolated fro the rat brain. The order of relative efficacy of the extracts in both experiments was as follows: C < D equal to or greater than A < B. Using EPR spectroscopy, the Kampo extracts were found to be oxidated by means of PbO2 and to become relatively stable radicals, which shows that they possess electro-donor properties. A relationship between their ability to inhibit peroxidation of LDL and liposomes and their ability to become a radical was found. The Kampo medicines which produced more radicals inhibited peroxidation of LDL and liposomes more effectively. By means of EPR spectroscopy, Kampo B was found to reduce the vitamin E radical. The results contribute to the understanding of the positive effects of Kampo extracts in the diseases in which a negative influence of free radicals is assumed.


[Local anesthetics. CVI. Preparation and effect of (+/-)-trans-[2-(piperidinomethyl) cyclopentyl] alkyl, alkoxy and halogen substituted esters of phenylcarbamic acid]. Within the framework of the research of potential local anaesthetics, five compounds from the group of (+/-)-trans-[2-(piperidino-methyl)cyclopentyl]alkyl, alkoxy and chloroalkyl esters of phenylcarbamic acid were prepared. The final compounds were obtained by addition of (+/-)-trans-2-(piperidinomethyl)cyclopentanol to the corresponding substituted phenylisocyanates. They were isolated in the form of salts with hydrochloric acid. Their structure was confirmed on the basis of interpretation of the results of UV and IR spectra. A pharmacological evaluation revealed that the final compounds were more effective than the employed standards cocaine and procaine, but they did not achieve the expected effect in comparison with the pattern compound pentacaine. The index of local-anaesthetic efficacy ranges from 1 to 8 in surface anaesthesia and 1 to 12 in infiltration anaesthesia. It is substantially lower even in comparison with analogical basic esters of alkoxyphenylcarbamic acid.


Molecular basis and allele specific screening of apolipoprotein CIISt. Michael. The DNA basis for a circulating nonfunctional variant apoCII, designated apoCIISt. Michael (CII-S) is a shifted reading frame. We developed two amplification primers to screen for CII-S. One contained wild type apoCII sequence and the other contained the sequence for CII-S. Allele-specific polymerase chain reaction (ASPCR) of the apoCII gene from the CII-S proband and her relatives showed unambiguous identification of heterozygous carriers of the CII-S protein. ASPCR may have general applicability in screening for DNA point mutations that do not alter a restriction site.


Observed relationship between ratios HDL-cholesterol/total cholesterol and apolipoprotein A1/apolipoprotein B. Epidemiological evidence suggests that the ratio HDL-cholesterol (HDL-C)/total cholesterol (TC) or apolipoprotein A1 (apo A1)/apolipoprotein B (apo B) are good indicators of coronary heart disease risk. In investigating the distribution of these ratios in the typical population served by our routine laboratory, we analysed the lipid results of 541 serum samples submitted over a 2-month period for TC, HDL-C, apo A1, and apo B. Good correlation was observed between HDL-C and apo A1 (r = 0.664), and between TC and apo B (r = 0.674). Surprisingly, the correlation between the ratios HDL-C/TC (range: 0.05-0.40) and apo A1/apo B (range: 0.27-3.71) was even higher (r = 0.822). Similar significant correlations were observed in 31 heterozygous and 20 homozygous familial hypercholesterolemic subjects, viz. the correlations between HDL-C/TC (ranges: 0.04-0.24 and 0.02-0.12, respectively) and apo A1/apo B (ranges: 0.47-1.84 and 0.15-1.12, respectively) were r = 0.951 and r = 0.972, respectively.


Role of cholesterol-accumulating macrophages on vascular smooth muscle cell proliferation. The proliferation of vascular smooth muscle cells (VSMC) was stimulated by co-incubation with macrophages. Further stimulation was observed when co-incubated macrophages were supplied with LDL or cholesterol. However, the stimulation of VSMC proliferation did not result from co-incubation with macrophages supplemented with acetylated LDL or delipidated LDL. The addition of anti-PDGF antibody partially abolished the stimulation of VSMC proliferation induced by co-incubation with macrophages supplemented with LDL or cholesterol. A high concentration of prostaglandin E2 inhibited the proliferation of VSMC stimulated by PDGF and plasma-derived serum when they were at the G0/G1 stage. However, the inhibitory effect of prostaglandin E2 on proliferation was not observed when cells were incubated with macrophages supplemented with LDL or cholesterol in spite of the promotion under these conditions of prostaglandin E2 production. These results suggest that cholesterol-accumulating macrophages may exert a regulatory effect on the proliferation of VSMC through the synthesis and secretion of platelet-derived growth factor (PDGF) and prostaglandin E2, besides foam-cell formation.


Eicosapentaenoic acid inhibits cholesteryl ester accumulation in rat peritoneal macrophages by decreasing the number of specific binding sites of acetyl LDL. We found that rat peritoneal macrophages bind acetyl low density lipoprotein (AcLDL) by a saturable and specific manner and accumulate a substantial amount of cholesteryl ester (CE) when incubated with AcLDL. In macrophages enriched with eicosapentaenoic acid (EPA) by the ingestion of highly purified eicosapentaenoic acid-ethyl ester (EPA-E), the accumulation of CE was significantly decreased in a dose-dependent manner. In addition, the contents of EPA and docosapentaenoic acid (DPA) in macrophage phospholipids were dose dependently and significantly increased by EPA-E feeding. In contrast, the contents of arachidonic acid (AA) and docosahexaenoic acid (DHA) were unchanged. Furthermore EPA-E ingestion significantly decreased the Bmax of the AcLDL receptor without affecting the Kd in rat peritoneal macrophages. In addition, specific proteolytic degradation of AcLDL was also dose dependently inhibited by EPA-E feeding, indicating that the number of AcLDL receptor was significantly decreased after EPA-E ingestion. These findings indicate that EPA-E feeding inhibited CE accumulation mainly by decreasing the AcLDL receptors in macrophages. We speculate that EPA inhibits foam cell formation and this inhibitory effect may partly account for its anti-atherogenic action.


Low-density lipoproteins in hyperalphalipoproteinemic heavy alcohol drinkers have reduced affinity for the low-density lipoprotein receptor. Heavy alcohol intake causes a marked inhibition of cholesteryl ester transfer protein (CETP) activity resulting in cholesterol ester enrichment of HDL. In this study we have characterized LDL of 35 chronic heavy alcohol drinkers with hyperalphalipoproteinemia to clarify the effect of alcohol on the metabolism of LDL. Serum concentrations of LDL-cholesterol and apolipoprotein B were normal, while the chemical composition of LDL was characterized by depletion of cholesteryl ester and enrichment of triglyceride. The LDL particles of the drinkers were significantly smaller in size than those of controls and had reduced affinity for LDL receptors of normal human fibroblasts. After cessation of alcohol, these abnormal characteristics returned toward normal along with elevation of CETP activity. These results suggest that heavy alcohol intake alters the compositions and particle size of LDL, consequently reducing their affinity for LDL receptors. This may be attributed, at least in part, to the reduction of CETP activity.