Hydrothermal Ethanol Flames in Co-Flow Jets.

Results on the autoignition and stabilization of ethanol hydrothermal fames in a Supercritical Water Oxidation (SCWO) reactor operating at constant pressure are reported. The flames are observed as luminous reaction zones occurring in supercritical water; i.e., water at conditions above its critical point (approximately 22 MPa and 374 °C). A co-flow injector is used to inject fuel (inner flow), comprising an aqueous solution ranging from 20 %-v to 50 %-v ethanol, and air (annular flow) into a reactor filled with supercritical water at approximately 24.3 MPa and 425 °C. Results show hydrothermal fames are autoignited and form diffusion flames which exhibit laminar and/or turbulent features depending upon flow conditions. Two orthogonal camera views are used; one providing a backlit shadowgraphic image of the co-flow jet and the other providing color images of the flame. In addition, spectroscopic measurements of flame emissions in the UV and visible spectrum are discussed.

The Salivary Mycobiome Contains 2 Ecologically Distinct Mycotypes.

A broad range of fungi has been detected in molecular surveys of the oral mycobiome. However, knowledge is still lacking on interindividual variability of these communities and the ecologic and clinical significance of oral fungal commensals. In this cross-sectional study, we use internal transcribed spacer 1 amplicon sequencing to evaluate the salivary mycobiome in 59 subjects, 36 of whom were scheduled to receive cancer chemotherapy. Analysis of the broad population structure of fungal communities in the whole cohort identified 2 well-demarcated genus-level community types (mycotypes), with Candida and Malassezia as the main taxa driving cluster partitioning. The Candida mycotype had lower diversity than the Malassezia mycotype and was positively correlated with cancer and steroid use in these subjects, smoking, caries, utilizing a removable prosthesis, and plaque index. Mycotypes were also associated with metabolically distinct bacteria indicative of divergent oral environments, with aciduric species enriched in the Candida mycotype and inflammophilic bacteria increased in the Malassezia mycotype. Similar to their fungal counterparts, coexisting bacterial communities associated with the Candida mycotype showed lower diversity than those associated with the Malassezia mycotype, suggesting that common environmental pressures affected bacteria and fungi. Mycotypes were also seen in an independent cohort of 24 subjects, in which cultivation revealed Malassezia as viable oral mycobiome members, although the low-abundance Malassezia sympodialis was the only Malassezia species recovered. There was a high degree of concordance between the molecular detection and cultivability of Candida, while cultivation showed low sensitivity for detection of the Malassezia mycotype. Overall, our work provides insights into the oral mycobiome landscape, revealing 2 community classes with apparently distinct ecologic constraints and specific associations with coexisting bacteria and clinical parameters. The utility of mycotypes as biomarkers for oral diseases warrants further study.

Platelet antibodies in immune thrombocytopenia.

Several tests to detect antibodies to platelets in patients with immune thrombocytopenia have been developed over the 40 years since it was first noted that this disorder was mediated by antiplatelet factors. Early tests were crude and not reproducible. A major landmark was the quantitation of IgG immunoglobulin on platelet membrane and the observation by several workers that marked increases of all classes of immunoglobulin occurred on platelets in patients with immune thrombocytopenia. Although at one time accepted as a diagnostic characteristic of autoimmune thrombocytopenia (AITP), the initial euphoria was tempered over the last decade by the realisation that elevation of platelet associated immunoproteins was not pathognomonic of this disorder and that raised levels were seen in several other disease processes, sometimes even when platelet counts were normal. The nature of these immunoproteins needs careful understanding. True platelet autoantibodies will manifest as increased platelet immunoproteins but not all such platelet proteins are platelet antibodies. There is speculation about the existence and the mode of activity of IgA and IgM immunoglobulins, both commonly found on platelets in AITP. It is sometimes almost inconceivable that the extremely large amounts of PAIgG could possibly represent true autoantibody. Immune complexes are found in plenty in these and other disorders in which thrombocytopenia manifest. In such situations it is likely that 'amplified' immune complexes are bound by Fc receptors, as may be found in viral mediated ITP or in septicaemic states. There is now sound evidence that several glycoprotein such as GP IIb/IIIa, GP Ib, GP V, found on platelet membrane, act as target antigen sites for the attachment of antibodies to platelets.(ABSTRACT TRUNCATED AT 250 WORDS)

Bcl-2 and c-myc expression, cell cycle kinetics and apoptosis during the progression of chronic myelogenous leukemia from diagnosis to blastic phase.

Chronic myelogenous leukemia (CML) has a progressive course but little is known about the biologic characteristics of disease progression. This study was designed to assess the changes in cell proliferative characteristics, apoptosis, the expression of the bcl-2 and c-myc genes between the time of initial diagnosis and entrance into the blastic phase of the disease. We observed that the rate of cell proliferation decreased and the cell death rate did not significantly change as the disease accelerated. The level of bcl-2 expression was significantly higher in accelerated/blastic phase cells than in the chronic phase cells in the population as a whole, however, the bcl-2 expression level did not change in blast cell subpopulation. c-myc Expression was significantly higher in the blast cell subpopulation of accelerated/blastic phase than in that of earlier phases of the disease. In conclusion, the characteristics of CML cells, namely proliferation rate, c-myc and bcl-2 change during the course of the disease. It is possible that the change in c-myc expression plays a causative role in evolution of the blastic phase from the chronic phase.

The effects of 13-cis retinoic acid and interferon-alpha in chronic myelogenous leukemia cells in vivo in patients.

The effects of the administration of a 3-day course of 13-cis retinoic acid in combination with interferon a [RA/IFN] on the leukemia cells was measured in vivo in 43 patients with chronic myelogenous leukemia. The administration of RA/IFN was associated with a significant fall in the white blood cell count of patients with chronic-phase disease and with a fall in the percentage S-phase cells in CML patients regardless of the stage of their leukemia. In two thirds of the patients studied the administration of RA/IFN was also associated with an increase in marrow apoptosis. The cytokine combination also suppressed bcl-2 and myc expression in a minority of patients and such expression appears to be associated with response to a treatment regimen which includes RA/IFN. These studies are the first to directly assess the effects of the combination of RA/IFN on chronic myelogenous leukemia cells in vivo in patients. These effects, if seen in other malignant diseases, could account for the therapeutic benefit which has been associated with the administration of this combination of biological agents to patients with malignant disease.

Phytohaemagglutinin as a molecular probe to study the membrane constituents of human X- and Y-bearing spermatozoa.

Phytohaemagglutinin (PHA) was employed to study the constituents of the cell membrane in X- and Y-bearing human spermatozoa. The presence of certain receptors for PHA was revealed in the equatorial region of about 50% of the spermatozoa. To determine the association of the equatorial staining by PHA with one of the subpopulations of spermatozoa a double staining technique was developed. In this method, the membrane was first stained with PHA employing the indirect immunofluorescence technique and then followed by the conventional quinacrine staining method to demonstrate the F-body. It was shown that spermatozoa which had receptors in the equatorial region for PHA were the ones which exhibited in the F-body. Staining of the membrane with PHA is a better method for the identification of human Y spermatozoa since it does not have the drawbacks of quinacrine staining.

Splenic B cell lymphoma with circulating villous lymphocytes: differential diagnosis of B cell leukaemias with large spleens.

The clinical, haematological, morphological and histological features of a series of 22 patients presenting with splenic lymphoma with circulating villous lymphocytes were assessed and compared with those of patients with other forms of chronic B cell leukaemia in an attempt to differentiate this condition from hairy cell leukaemia, prolymphocytic leukaemia, and chronic lymphocytic leukaemia, with which this condition has many features in common. The disease was twice as common in men than in women, with a mean (SD) age at diagnosis of 72 (9) years, and the most consistent presenting feature was massive enlargement of the spleen, which showed white and red pulp disease with a plasmacytic component. Small monoclonal bands were found in 60% of cases.

Diagnosis of alpha-thalassemia trait from Coulter Counter 'S' indices.

A number of patients of Mediterranean and Asian origins were found to have unexplained microcytic hypochromic red blood cells. Iron deficiency and beta-thalassaemia trait were both satisfactorily excluded in all of them. The haematological indices of these patients, obtained on a Coulter Model 'S' Counter, were found to be very similar to those seen in obligatory heterozygotes for alpha-thalassaemia. It is postulated that these patients were also carriers for alpha-thalassaemia. Subsequent investigation of some of these patients showed the characteristically reduced rates of alpha-chain synthesis seen in this condition. The discriminant function of England and Fraser (1973) may be of help in diagnosing this state. alpha-Thalassaemia trait should be considered in all patients of 'high-risk' ethnic origins with a blood picture suggestive of beta-thalassaemia trait but in whom the levels of Hb A2 and Hb F are within normal limits.

Hereditary angioedema and thyroid autoimmunity.

Sera from 91 patients with hereditary angioedema were screened for thyroid antibodies. The results for the 77 patients more than 17 years old were compared with previously published data for the prevalence of thyroid disease in a large community (Whickham). Of the female patients with hereditary angioedema, the prevalence of thyroglobulin antibodies (TGA) was 14.0%, higher than the expected 3% (p less than 0.001). The prevalence of thyroid microsomal antibodies (TMA) was 20%, also higher than the expected 7.6% (p less than 0.01). The age distributions of the females in both groups differed: in the group with hereditary angioedema there was a greater proportion of younger patients which should have resulted in a lower prevalence of thyroid antibodies. Adjusting for related patients with hereditary angioedema, there was still an increased prevalence of TGA (p less than 0.01) and TMA (p less than 0.01).

Adult T-cell lymphoma/leukemia in a Caribbean patient: cytogenetic, immunologic, and ultrastructural findings.

Cytogenetic findings on immunologically and morphologically characterized leukemic cells from a Caribbean patient with adult T-cell lymphoma/leukemia (ATLL) (HTLV+) are reported. Marker studies on peripheral blood lymphoid cells showed a mature postthymic phenotype: TdT-, OKT3+, OKT4+, OKT6-, OKT8-, 3A1-, anti-HLA-DR-. Light and electron microscopic analysis revealed a great cellular pleomorphism with respect to nuclear features. Three main types of leukemic cell were observed: typical multilobed ATLL lymphocytes, Sézary's syndrome (SS) cells, and cells intermediate between those two. Chromosome studies on PHA-stimulated cultures revealed three clones. The predominant clone was hyperdiploid; significant abnormalities were 1q+, 14q+, and 6q- (breakpoint q21), which are known to occur in lymphoid malignancies, together with trisomy 7q and i(17q), which have been reported previously in Japanese ATLL and the small variant of SS, respectively. The 14q+ marker was t(11;14)(q13;q22-24). The incidence of 6q-, trisomy 7q, and i(17q) varied within the main clone, and it is speculated that these chromosome abnormalities might be related to the variation observed in the cell types of this patient. Two minor clones had 6q- (breakpoint q25) and 13q+ markers, respectively. It was not possible to unequivocally establish the relationship between these three clones. The chromosomal, morphologic, and immunologic findings in this case support a close relationship between ATLL in Japan and in the Caribbean basin, as well as between the proliferating cells of ATLL and SS.

The Splenic White Pulp in Chronic Lymphocytic Leukaemia: A Microenvironment Associated with CR2 (CD21) Expression, Cell Transformation and Proliferation.

Splenectomy specimens from 8 cases of chronic lymphocytic leukaemia (CLL) were examined. The infiltrate in the red pulp consisted predominantly of small lymphocytes. In contrast the predominant cells in the white pulp were pro-lymphocytes and para-immunoblasts. The Ki67 marker, which identifies cells in growth phase, was also concentrated among the transformed cells in the white pulp. Staining with monoclonal antibodies for CD21, which identifies the CR2 receptor, showed that the cells in the white pulp were strongly positive in contrast to those in the red pulp which were weak or negative. These findings suggest that factors present in the white pulp promote transformation and proliferation of CLL cells.

Chronic myeloid leukemia following radioactive iodine (131)I therapy of thyroid cancer.

We describe a 51 years old female patient who developed Ph(1) positive chronic myeloid leukemia 7 years following radioactive iodine therapy for follicular carcinoma of thyroid. Until now, only two patients have been reported to have developed CML after this kind of therapy. The underlying mechanisms are discussed and the need to study such patients at cytogenetic, molecular biologic and cell kinetic levels is stressed.

Accuracy of immunohistochemistry and flow cytometry in estimating the proportion of leukemic cells in S phase in chronic myeloid leukemia patients.

We compared the proportion of S phase cells in bone marrow and peripheral blood samples obtained from 17 patients with chronic myeloid leukemia (CML). Before sampling all patients received a one hour i.v. infusion of iododeoxyuridine (IdUrd). The proportion of S phase cells was studied by immunohistochemistry (IHC) in bone marrow biopsies, and by flow cytometry (FCM) in bone marrow aspirates and peripheral blood samples. The IdUrd labelling index (LI) in bone marrow biopsy sections (27.5 +/- 1.8%) was significantly higher than the proportion of IdUrd labelled cells in bone marrow aspirate (15.1 +/- 2.0%). The percentage of S phase cells in peripheral blood was approximately the same as that in the aspirate (12.4 +/- 1.3%) and was correlated with that of bone marrow aspirate indicating a high degree of the aspirate dilution by peripheral blood. It is likely that the differences in % S phase cells in the aspirate and the biopsy result from this dilution. Estimates of the % S phase cells in the peripheral blood study by IHC and FCM were essentially the same. Samples labelled for one hour in vitro resulted in 1.5 fold higher LI than the same samples labelled in vivo. We conclude that estimates of the 8% S phase cells in the bone marrow of patients with CML should be made by infusing patients with IdUrd or BrdUrd with immunohistochemical evaluation of a marrow biopsy. Additionally in vitro labelling is not reflective of the percent S phase cells in vivo in patients.

A new IVIgG preparation (IVIgG pH 4.25): preliminary results for the treatment of immune thrombocytopenia.

Fifteen patients comprising ten with chronic idiopathic autoimmune thrombocytopenic purpura, three with acute autoimmune thrombocytopenic purpura, and two with secondary chronic autoimmune thrombocytopenic purpura were treated. All patients received 5-day courses of intravenous immunoglobulin (pH 4.25) at a dose of 400 mg kg body weight daily and five patients received maintenance treatment with double dose single infusions. Responses were seen in 12 patients which were in general of brief duration although one patient had a sustained remission. Five patients were treated with maintenance therapy of 800 mg/kg given at approximately 3-weekly intervals and maintained satisfactory platelet counts.

Monoclonal anti-idiotypic antibody to progesterone with internal image properties.

An auto-anti-idiotypic approach was used to generate mouse monoclonal anti-idiotypic antibody E9F11F6 (Ab2) to progesterone. A conjugate of 4-pregnane 3,20 dione and bovine serum albumin was used as the immunogen. E9F11F6 bound to a rabbit anti-progesterone antibody in a linear concentration-dependent manner. It inhibited the binding of [3H]progesterone to a monoclonal anti-progesterone antibody, E9D5 (Ab1); this inhibition was concentration dependent. Results from chase experiments showed that this inhibitory activity of Ab2 was not due to steric hindrance but was a result of direct binding to the ligand combining site. Indirect immunofluorescence studies also showed that Ab2 and progesterone bound to similar binding sites on Ab1. Overall, the results suggest that E9F11F6 contains an internal image of at least part of the progesterone molecule, and that it can be classified as an Ab2 beta antibody. This anti-idiotypic antibody may be of value in further studies of endocrine and reproductive physiology.

Immune response to syngeneic spermatozoa & its effect on target organs in mice.

Two groups of adult Swiss mice were immunised with washed syngeneic spermatozoa without any adjuvant for a period of two months or four months respectively. The presence of antibodies to spermatozoa was measured by micro sperm-agglutination and micro sperm-immobilization tests. The development of cell mediated immune response (CMIR) was measured by leucocyte migration inhibition test (LMIT) using spermatozoal antigens solubilized by 3M KCl, Nonidet P-40 or by subjecting the cells to ultrasonication. SDS-PAGE analysis of these proteins indicated that extraction of spermatozoa with 3 M KCl was a better method for solubilization of antigens present on sperm membrane. Almost all immunized mice had varying titers of sperm agglutinating antibodies. Nearly 40-50 per cent of the mice had a titre of 1:128 in both groups whereas only 33 per cent had sperm immobilizing antibodies. CMIR, as assessed by LMIT, was detected in immunized mice. However, this had not resulted in the infiltration of immune cells into the target organs perhaps due to the lower magnitude of immune response.

A simple and reproducible method for separating Y-bearing spermatozoa from human semen.

PIP: A simple and reproducible method for separating Y-bearing spermatozoa from human semen is reported. Semen samples with 60-80 million count and 50% motility were obtained from healthy human donors. Ficoll-400 and sodium metrizoate density gradient were used for separation. Quinacrine hydrochloride was used for staining, and Olympus fluorescent microscopy was used for the detection of F body present in the spermatozoa bearing Y-chromosomes. The data indicate that the percentage of Y-bearing spermatozoa in normal human semen is about 5-6% less than what is expected. The observations agree with those of Barlow and Vosa that with the optical attenuation coefficient being sufficiently great, the chromosomes lying deep in the head may go undetected. A mean value of 45.22 was obtained for Y-bearing spermatozoa and the sediment contained about 62.9 Y-bearing spermatozoa. The results thus suggest that it is possible to separate Y-bearing spermatozoa from semen using Ficoll-socium metrizoate density gradient.^ieng

Prognostic significance of myeloperoxidase containing blast cells in acute myelogenous leukaemia.

Fifty three newly diagnosed patients of de novo acute myelogenous leukaemia (AML) received treatment consisting of remission induction with daunorubicin 60 mg/m2 on day one and continuous infusion of cytosine arabinoside 200 mg/m2/day over 24 h from day one to 7. Thereafter patients in complete remission received consolidation chemotherapy with two identical courses. Complete remission (CR) could be achieved in 40 patients (75.5%). Seven patients (13.2%) died with complications during aplasia phase following remission induction therapy while six patients (11.3%) had resistant disease. Twenty seven patients (67.5%) developed relapse while eight patients (15.1%) continue to remain in complete remission ranging from 51 to 68 months (median 62.5). The projected event free survival and disease free survival at 60 months is 15 per cent (SE + 11.9%) and 21 per cent (+6%) respectively. Evaluation of the prognostic significance of pretherapy characteristics showed that infection at presentation and low number of myeloperoxidase (MPO) containing blasts affected the achievement of complete remission adversely on univariate analysis. Similarly age at diagnosis, of more than 45 yr, total leucocyte count of 50,000/cumm or more and low number of MPO containing blasts affected the remission duration (disease free survival) adversely on univariate analysis. On multivariate analysis, MPO positivity of blast cells, remained the only significant independent characteristic. High MPO positivity affected the remission duration favourably (P < 0.01). Patients with high MPO positivity also achieved CR with one induction cycle in 32 out of 40 instances while only 2 out of 5 patients with low MPO positivity, achieved CR with one chemotherapy cycle (P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Immunomodulation of the mother during pregnancy.

The concept that the immune responsiveness of the mother is reduced during pregnancy arose from studies which appeared to show that immune response to certain antigens is reduced during pregnancy (1, 2). Various substances claimed to have immunosuppressive or immunomodulating effect include alpha fetoprotein, placental proteins, early pregnancy factor (EPF), human chorionic gonadotropin (HCG), corticosteroids, estrogens, androgens and progesterone (2). To summarise a body of literature, there is very little change in the immune competence of the mother during pregnancy. This makes sense, as generalized immunosuppression would be a risky way to ensure the survival of the fetus. Immune enhancement and subsequent immunomodulation of the mother is likely to be the mechanism operative during pregnancy. It is conceivable that the overall immune response in pregnancy could be the net result of an interplay of various interactions that may be operating to ensure non-rejection of the antigenically alien fetus while at the same time preventing a state of excessive immunosuppression. Such a dynamic homeostatic mechanism appears to be important for the successful completion of pregnancy.

Production and characterisation of a monoclonal antibody to progesterone and its effect on fertility.

A monoclonal antibody reacting with progesterone has been raised by fusion of mouse myeloma cells (SP20) and splenocytes of BALB/c mice hyperimmunized with 4-pregnane 3,20 dione conjugated to bovine serum albumin. Association constant of this antibody for binding with progesterone was 0.22 x 10(9) l/mole. The antibody was highly specific for progesterone. A single ip injection of this antibody brought about an antifertility effect which is influenced by genotype. Antibody treatment brought about a significant decrease in the fetal weight and a slight decrease in the plasma progesterone levels. The antifertility effect could be reversed only up to day 3 by exogenous administration of progesterone.