Activation-induced cell death in human peripheral blood lymphocytes after stimulation with silicate in vitro

Silica and related substances such as silicate have been proven to possess "adjuvant effects". We have previously reported a finding of polyclonal human T cell activation induced by silicate as a superantigen in vitro. In this study, we observed activation-induced cell death in human lymphocytes after stimulation with chrysotile, a kind of silicate. Apoptotic cells were detected flow cytometrically using the TUNEL assay, and the maximum appearance of TUNEL positive cells occurred on day 4 of incubation. Simultaneously the manifestation of small-sized cells in the specimens increased implying apoptosis. Fas expression on lymphocytes increased to day 3 of incubation with chrysotile, and then spontaneously decreased on day 4 when remarkable apoptosis could be detected. Based on these results it is conceivable that activation-induced cell death occurred through Fas-Fas ligand interaction in lymphocytes after stimulation with silicate in a concentration with which no acute cytotoxicity has been detected. Whether and how the repeated apoptosis in definite clones of lymphocytes causes the induction of sFas synthesis need clarification.     

Flow-cytometric analysis of peripheral blood lymphocytes in patients with chronic idiopathic neutropenia of adults

Blood traces tortuous paths of flow through the heart. We postulate that momentum changes associated with direction changes optimize dynamic coupling between ventricular and atrial function, particularly on exercise. Traces of pulsed Doppler mitral flow and M-mode long-axis mitral ring movement were recorded before and during exercise, increased in 25-W steps to strenuous levels (146 +/- 30 W), in 16 healthy volunteers, aged 38 +/- 10 years. R-R intervals fell from 821 +/- 151 to 437 +/- 51 ms, and diastole from 458 +/- 134 to 169 +/- 33 ms. Peak mitral flow velocities rose from 0.68 +/- 0.17 to 1.27 +/- 0.16 m/s, and mitral valve ring displacements from 13.8 +/- 3.3 to 19.3 +/- 3.4 mm. Biphasic diastolic curves of flow and movement became monophasic as R-R fell below 500 ms, with atrial systole apparently coming to coincide with elastic ventricular recoil to give a single elevated peak of mitral flow. The increased slope and amplitude of Doppler curves indicate increased rates of change of momentum, which imply enhanced inertial forces. The illustrated patterns of flow and movement on exercise accord with the postulated "dynamic" mode of function, in which forces between atria, ventricles, and passing blood masses become tightly coupled to achieve a sling-like redirection of momentum through tortuous paths of flow, but more extensive data are needed to adequately model and quantify inertial force exchanges of the exercising heart.     

Antitumor killer lymphocytes in the peripheral blood of a patient with transitional cell carcinoma of the bladder

Despite the trend in the use of electromechanical left ventricular assist devices (LVAD), the highly compact, long lasting, and endurance characteristics of the new pneumatic system has its place in the treatment of end-stage chronic heart failure. The ProCor Model 1 LVAD has an implantable pump which is small in size and has a double pusher plate design. The portable power pack is small, too. It is easy to carry, and it is synchronized to the ECG to pump during the patient's diastolic period. Constant driving conditions are set as follows: frequency rate, 60 bpm; systolic percentage, 25 (250 ms); dP/dt, 1,800 (45 ms) mm Hg.s; and driving pressure, 250 mm Hg to have a basic 3.500 L/min pump output for prolonged circulatory assistance. The 6 mm internal diameter (ID) percutaneous multipurpose pneumatic tube contains within its wall both a spiral of MP35N alloy wire connected to the myocardial lead for ECG sensing and a spiral serving as mass ready to be used for cardiac pacing with an external pacemaker when necessary. The aortic porcine bioprosthesis maintains the aortic root and the sinus ridge.     

Tropism of human herpesvirus 8 for peripheral blood lymphocytes in patients with Castleman's disease

Multicentric Castleman's disease (MCD), also called multicentric angiofollicular lymphoid hyperplasia, is a systemic lymphoproliferative disorder causing fever, lymphadenopathy and splenomegaly. Recently, Kaposi's sarcoma-associated herpesvirus/human herpesvirus 8 (KSHV/HHV-8) DNA sequences have been detected in cases of MCD. We examined HHV-8 DNA sequences in the peripheral blood mononuclear cells (PBMCs) of two HIV-negative patients with MCD and in PBMCs and the lymph node of a HIV-negative patient with localized Castleman's disease (LCD) by the polymerase chain reaction. The novel sequences were detected in all DNA samples. Furthermore, the sequences were detected in only the CD19+ B-lymphocyte fraction of the patient with LCD as previously reported. However, the sequences were detected in CD19+ B-lymphocyte and CD2+ T-lymphocyte fractions of two patients with MCD. These results suggest that HHV-8 has tropisms for both B lymphocytes and T lymphocytes in Castleman's disease.     

Functional analysis of peripheral blood B cells in patients with X-linked agammaglobulinemia

X-linked agammaglobulinemia (XLA) is a primary immunodeficiency disease caused by mutations of Bruton tyrosine kinase (Btk); Btk plays an essential role in the development of mature B cells. However, small numbers of B cells ("leaky B cells") are present in the peripheral blood of most XLA patients. In this study, we analyzed the function of these leaky B cells obtained from XLA patients. Enough numbers of B cells were available for analysis from five of nine XLA patients originally screened. Sequence analysis revealed missense mutations of Btk in four of the five XLA patients. No mutation was found in the coding region of Btk in one patient. Western blotting and/or flow cytometric analysis failed to detect Btk protein in all five patients. B cells isolated from peripheral blood of these XLA patients were CD5-, CD20+, CD19+, and CD21-. If stimulated with anti-CD40 and IL-4, XLA B cells proliferated normally and produced significant amounts of IgE. Anti-CD40 stimulation of XLA B cells resulted in normal expression of CD23. In addition, three of the five XLA patients studied were immunized with bacteriophage phiX174 and produced low but detectable levels of antiphage-specific Ab. Similarly, X-linked immunodeficiency mice, which carry a missense mutation in Btk, produced substantial amounts of antiphage Ab. These results indicate that CD40 signaling is intact in B cells lacking demonstrable Btk, and that leaky B cells in XLA patients can proliferate, undergo isotype switching, and differentiate into specific Ab-producing cells.     

Defective natural killer cell activity of peripheral blood lymphocytes correlates with the degree of neutropenia in patients with chronic idiopathic neutropenia of adults

Natural killer cell activity (Nka) of peripheral blood mononuclear cells (PBMCs) against K562 cell targets was assessed in 66 patients with chronic idiopathic neutropenia of adults (CINA) using the 16-h 51Cr-release assay. It was found that CINA patients exhibited significantly lower Nkr than normal subjects, which strongly correlated with the degree of neutropenia and the numbers of circulating neutrophils. Patients' NKa was increased by recombinant human interleukin-2 (rhIL-2) or recombinant human interferon-alpha (rhIFN-alpha), but the values obtained did not reach the respective NKa values found in normals. However, percentages of cytokine-induced rises of NKa did not differ statistically between patients and normal subjects. No serum inhibitors of NKa were demonstrated in our patients. CINA patients had low numbers of circulating NK cells as defined by the expression of NK-cell-related surface markers CD16, CD56, and CD57. CD16+ and CD56+, but not CD57+, cells correlated with the values of baseline NKa. The numbers of all these cell subsets correlated with the degree of neutropenia and the numbers of circulating neutrophils. Using CD56+-enriched PBL suspensions, it was shown that patients' NK cells displayed normal tumor cell binding capacity and produced in vitro normal amounts of natural killer cytotoxic factor(s) against K562 cell targets upon activation with rhIFN-alpha. Finally, percentages of perforin-expressing and granzyme B-expressing CD16+ cells did not differ statistically between patients and normal controls. Based on all these observations, we concluded that CINA patients display low NKa probably because they have low numbers of circulating NK cells. No functional abnormalities of NK cells were demonstrated. The cause and the underlying mechanisms leading to NK-cell depletion in these patients remain to be clarified.     

Telomerase activation in normal B lymphocytes and non-Hodgkin's lymphomas

Activation of telomerase seems to be a prerequisite for immortalization and is found in permanent cell lines and most malignant tumors. Normal somatic cells are generally telomerase negative, except for bone marrow stem cells. Weak activity is also present in peripheral blood cells. In the present study strong telomerase activity was demonstrated in vivo in normal mature cells of the immune system, as well as in malignant lymphomas. Benign lymph nodes had lower telomerase activity than benign tonsils, which exhibited intermediate to high activity comparable with findings in malignant lymphomas. In benign tonsils the activity seemed to be restricted to germinal center B cells. In benign lymphoid tissues telomerase activity correlated with B-cell numbers and cell proliferation, but this was not observed in the lymphoma group. High-grade lymphomas exhibited higher levels of telomerase compared with low-grade cases. The data showed that in vivo activation of telomerase is a characteristic feature of germinal center B cells. Different signals for activation of telomerase are likely to exist, one of them being immune stimulation. The data suggest that telomerase activity in malignant lymphomas can be explained by an "induction and retention" model, ie, transformation occurs in a normal, mature B cell with reactivated telomerase, which is retained in the neoplastic clone.     

Kinetics of cell division in peripheral blood lymphocytes of stainless steel welders]

The management of fracture within previously shortened extremity is presented. Three patients with femoral fracture and one with crural fracture were treated. All fractures were displaced. Modified Ilizarov ring distractor was used to stabilize the fracture within 3 days from injury. Distraction was commenced on third postoperative day at speed of 0.75-2 mm/h with progressive reduction of fragments and alignment of mechanical axis of the limb. Lengthening required to equalize extremities was achieved in all cases. The author claims fracture within previously shortened extremity to be one more indication for external osteosynthesis as a method of choice.     

Androstenedione conversion in human peripheral blood lymphocytes

Indigenous Indian groups comprise approximately 20% of Ecuador's population, the third largest percentage in all of Central or South America, yet immunogenetic data on these groups are lacking in the literature. In the course of population migration studies, sera collected from 65 Ecuadorians living in the northern province of Esmeraldas were typed for six GM and two KM markers. The study population consisted of 47 Cayapa Indians and 18 blacks of African origin, descendants of slaves imported into the area during the seventeenth century. The Cayapa demonstrated three GM phenotypes, two of which are common to other South American Indian tribes. The frequency of KM1 positive Cayapa Indians (63%) is similar to other South American Indian tribes, but is significantly greater than the Huaorani of eastern Ecuador (2%), the only other Ecuadorian Indian group for whom limited immunoglobulin allotype data are available (chi 2 = 35.8, P < 0.0001).     

Allogeneic peripheral blood stem cell transplantation in 30 patients with hematologic disorders

Thirty patients (median age of 32 years; range, 6-61) with hematologic disorders received unmanipulated peripheral blood stem cell transplants from HLA-matched or one-antigen-mismatched related donors following myeloablative therapy for acute lymphoblastic leukemia (7), acute myelogenous leukemia (6), chronic myelogenous leukemia (8), myelodysplastic syndrome (3), or other disorders (6). Granulocyte colony stimulating factor (G-CSF) mobilized peripheral blood stem cells were collected from donors in 1 to 3 aphereses. The apheresis products contained mean counts of 11.3 x 10(8) (range, 3.8-17.2) nucleated cells/kg and 6.7 x 10(6) (range, 1.3-16.7) CD34+ cells/kg. Graft-versus-host-disease (GVHD) prophylaxis consisted of cyclosporin A plus methotrexate, or FK506 plus methotrexate. All patients received G-CSF following their transplant. Although 1 patient died of pneumonia 6 days after transplantation, the others demonstrated rapid engraftment. Median days to recovery to 500/microliter neutrophils and 20,000/microliter platelets were 13 (range, 8-21) and 14 (range, 1-23) days, respectively. The incidence of acute GVHD grade II-IV was 33%; chronic GVHD developed in 57% of the assessable patients. There were no episodes of graft failure or rejection. Nineteen patients (63%) were alive and in complete remission from 147 to 839 days following their transplant (median follow-up of 560 days). Further follow-up study will be required to assess the incidence of chronic GVHD and graft-versus-leukemia (GVL) effects.     

Expression of CTLA-4 molecule in peripheral blood T lymphocytes from patients with systemic lupus erythematosus

The purpose of this study was to compare two planning techniques for the delivery of radical localized postoperative radiotherapy in the treatment of high grade astrocytomas. Conventional orthogonal plain film planning (CONP) was compared with CT planning (CTP) in terms of the size of the target volumes treated, the amount of normal brain irradiated and the accuracy of localization. Twenty consecutive adults with high grade astrocytomas, who were treated with radiotherapy between March and October 1996, were planned with CONP and CTP, using postoperative, contrast-enhanced CT scans to define the tumour volume. The planning target areas, volumes and the 100%, 80% and 50% isodose areas produced using the two planning methods were measured and compared using Student's paired t-test. The target volume length was also measured and compared as an independent factor. The difference between entry points of the central axis of the lateral fields was noted. Nineteen of 20 patients had a reduction in planning target volumes using CTP compared with CONP. The difference between the mean volumes was clinically relevant, with 288 cm3 representing a 25% reduction, and statistically significant at the P<0.001 level. The planning target volumes were reduced in 18/20 patients (mean 24 cm2, 23%, P<0.001). Similarly, there were highly significant reductions in the 100%, 80% and 50% isodose areas. The target volume lengths were not found to be significantly different. When considering the accuracy of localization, the entry point of the CONP lateral field deviated by a mean distance of 1.6 cm relative to CTP (superiorinferior 1.3 cm; anteriorposterior 0.8 cm; range 04.1 cm). In two patients, this would have led to a geographical miss of macroscopic disease. In patients with high grade astrocytomas, CTP is preferred to conventional planning. It leads to appreciable reductions in the size of the planning target areas and volumes receiving radical doses of radiation, it significantly reduces the amount of normal brain tissue being irradiated and is more accurate in terms of tumour localization. These differences are likely to lead to a reduction in treatment morbidity. We recommend CTP for all patients receiving radical radiotherapy.     

Alteration of signal-transducing molecules in tumor-infiltrating lymphocytes and peripheral blood T lymphocytes from human colorectal carcinoma patients

An analytical method for the rapid isolation and recovery of the homologous series of 2-aminoethanols, a class of organic compounds of importance to wood preservative treatment, is successfully developed. The method is applied to an aqueous solution of copper amine (copper[II] hydroxide complexed monoethanolamine) and to copper-amine-treated sawdust. The method incorporates a gas chromatograph-ion-trap mass spectrometer. A discussion of the secondary equilibrium effects involved when ionizable analytes are extracted from an aqueous phase with respect to organic bases is presented. Using 2-propanol as the extractant coupled to a salt-saturated aqueous phase results in recoveries of 63% for 2-aminoethanol, 51% for N,N-dimethyl-2-aminoethanol, and 56% for N-methyl-2-aminoethanol for a single liquid-liquid extraction. The choice of 2,2,2-trifluoroethanol as an internal standard is found to be quite suitable. A comparison of the precision and accuracy for an external versus an internal mode of instrument calibration demonstrates that the internal standard mode is preferable for this manual injection.     

Expression of human placental cell surface antigens on peripheral blood lymphocytes and lymphoblastoid cell lines

The expression of human placental cell surface antigens was examined in cells of lymphoid origin, including peripheral blood lymphocytes and cultured lymphoblastoid cells of bone marrow or thymus derivation. A select group of this defined set of surface antigens was detected on all three cell preparations. The most remarkable observation was the conspicuous absence of three subunits previously demonstrated to be present on all human cell surfaces examined to date. Antiserum directed against several placental components prevents adhesion and spreading of cells which grow attached to surfaces. These results suggest a role for these three glycoproteins in mediating cellular adhesion.     

Calcineurin protein phosphatase activity in peripheral blood lymphocytes

The protein phosphatase activity of peripheral blood T lymphocytes (PBLs) was examined to quantify the contribution of calcineurin and other members of the family of serine/threonine protein phosphatases. Using selective phosphatase inhibitors, the fractional phosphatase activities of calcineurin, protein phosphatases 1 (PP1), 2A (PP2A), and 2C (PP2C) were determined. Okadaic acid was used to inhibit the activity of both PP1 and PP2A while cyclosporin A/cyclophilin or trifluoperazine were used as a specific inhibitors of the calmodulin-dependent phosphatase calcineurin. Using a [32P]labeled 19-residue phosphopeptide substrate, RII peptide, it was found that PP1 and PP2A comprise the majority of the total phosphatase activity in PBLs with okadaic acid inhibiting 80% of the phosphatase activity. The remaining 20% of the phosphatase activity can be attributed primarily to calcineurin since it was Ca2+ dependent, sensitive to inhibition by the calmodulin antagonist trifluoperazine, and inhibited by the complex of cyclosporin A (CsA) and cyclophilin. These results indicate that PBL extracts contain little PP2C activity. In addition, PBLs treated with CsA had measurably lower calcineurin activity in cell lysates. The measurement of calcineurin activity may provide a useful means of assessing the extent of immunosuppression during drug therapy.     

Cytotoxic activity of platelets and peripheral blood mononuclear cells from oncology patients and healthy donors

Testicular germ cell tumour is said to be a model for curable neoplasm. However, the prognosis of primary extragonadal germ cell tumour does not appear to be as promising. Though similar in histology, the biology of primary extragonadal germ cell tumour is different as exemplified by the patients in this review. Eight patients with primary mediastinal germ cell tumours were treated with intensive cisplatin-based chemotherapy. All, except one, had non-seminomatous components. The poor prognosis of mediastinal germ cell tumour is due to a combination of poor treatment results with the cisplatin-based regimen and the development of non-germ cell and haematological malignancies.     

Impaired blastogenic response of lymphocytes from synovial fluid and peripheral blood of patients with rheumatoid arthritis

Synovial fluid lymphocytes from patients with rheumatoid arthritis demonstrated a markedly diminished blastogenic response to both phytohemagglutinin and pokeweed mitogens, when compared to normal peripheral blood lymphocytes. The blastogenic response to rheumatoid peripheral blood lymphocytes to both mitogens was also depressed, when compared to the response of normal lymphocytes, but the difference was less marked and was within limits which could be accounted for by recent salicylate therapy. Lymphocytes of both peripheral blood and synovial fluid of rheumatoid patients showed a delayed response to PHA (five days to achieve maximum thymidine incorporation vs four days for normals).     

Phenotyping of peripheral blood lymphocytes in adult coeliac disease

OBJECTIVE: To evaluate nerve fiber density in vestibular specimens from women operated upon for vulvar vestibulitis. METHODS: Forty-seven women with vulvar vestibulitis syndrome underwent modified posterior vestibulectomies. Vestibular specimens were analyzed after being stained for S-100 neural tissue protein. Women were followed up for 2 years. RESULTS: In specimens from 44 of 47 patients, the densities and numbers of nerve fibers per square unit in the preparations were greater than those in specimens from six control women. In the patients, a statistically significant linear correlation was found between inflammation and nerve bundle density in the preparations (Spearman rank correlation coefficient rs=.41; P=.005). There were no signs of infectious etiology in any preparation. No or slight postoperative dyspareunia was reported by 38 of 42 women after 6 months, 36 of 39 after 12 months, and 26 of 28 after 24 months. CONCLUSION: Vestibular neural hyperplasia may provide a morphologic explanation of the pain in vulvar vestibulitis syndrome.     

Spontaneous chromosome aberrations in the lymphocytes of human peripheral blood]

The authors carried out cytogenetic studies on lymphocytes from human peripheral blood of 105 persons--49 women and 56 men in order to examine spontaneous frequency of the structural chromosomal abberations in the somatic cells. There were 191 cells with abberations (1,17%), out of 16,267 analyzed metaphasic plates. They found 0,39% of chromatid fragments, 0,71% of chromosomal fragments, 0,06% dicentricets, 0,01% of symetric and asymetric chromatid exchanges and 0,006% of rings. There were no statistically significant differences in the discovery of the spontaneous abberations in the examined persons at various age and sex.