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At least 343 records · Page 19Linked to original sources
PubMed1987

Suppression of growth of PHA-stimulated human lymphocytes by a novel steroid (RU 28 362) lacking the 21-OH group.

RU 28 362 is a novel synthetic steroid (lacking the 21-OH group) which reacts exclusively with the glucocorticoid receptor. It is therefore chemically different from the other natural and synthetic glucocorticoids in common use which also bind to a lesser extent to receptors for other classes of steroids. Aspects of the immunosuppressive effect of RU 28 362 were studied by comparing its suppressive effects on PHA-stimulated growth of human lymphocytes with those of hydrocortisone, betamethasone and the inactive analogue, 21-deoxyhydrocortisone. RU 28 362 was more potent than either hydrocortisone or betamethasone, but all three glucocorticoids had a parallel dose-response curve in suppression of the increase in cell volume that occurs with activation during the first day in culture. In studies on the time of appearance and density of IL-2 receptors during the first 3 days in culture, RU 28 362 was also somewhat more inhibitory and had a steeper dose-response curve than hydrocortisone or betamethasone. RU 28 362 was much more inhibitory and had a steeper dose-response curve than either hydrocortisone or betamethasone in inhibiting [3H]-TdR incorporation by 3-day PHA stimulated cultures. 21-deoxyhydrocortisone did not inhibit any aspect of PHA-stimulated lymphocyte growth.

Androstanols↗
PubMed1984

Morphological details of the projection from the presubiculum to the entorhinal area as shown with the novel PHA-L immunohistochemical tracing method in the rat.

Iontophoretic injections of the lectin, phaseolus vulgaris leucoagglutinin (PHA-L) were made into the presubiculum of rats. The anterogradely transported lectin was visualized by using an anti-PHA-L antibody in combination with immunohistochemistry. The PHA-L tracing method revealed morphological details of the projection of the presubiculum to the ipsi- and contralateral medial entorhinal area usually not seen with other anterograde transport techniques. Fine varicose fibers form a dense terminal plexus in the deep parts of layer III. In layer II and deep layer I, the fibers form column-like axonal bundles, terminating in patches in the deep part of layer I. Some fibers reach the outer three layers of the entorhinal area (EA) from collaterals of axons running in the molecular layer, while a majority enter from the deep layers.

Animals↗
PubMed1988

Retrograde transport of the lectin Phaseolus vulgaris leucoagglutinin (PHA-L) by rat spinal motoneurons.

The lectin Phaseolus vulgaris leucoagglutinin (PHA-L) has been used primarily as an anterograde transport tracer in the CNS. We present evidence of PHA-L retrograde transport by rat spinal motoneurons after injection into the triceps brachii. Labelled motoneurons were localized in specific and well-defined neuron pools in the ventral horn. Primary afferent labelling was not seen in the spinal gray matter. Dorsal rhizotomy did not eliminate or decrease motoneuron labelling. The retrograde transport rate was about 8 mm/day. PHA-L can clearly undergo retrograde, as well as anterograde, transport.

Animals↗
PubMed1990

Topographical distribution of septohippocampal projections demonstrated by the PHA-L immunohistochemical method in rats.

The anterograde tracer, Phaseolus vulgaris-leucoagglutinin (PHA-L) was injected into the medial septum-diagonal band complex of the rat. When PHA-L was injected into the rostral part of the medial septum, labeled axon terminals were distributed largely in the dorsal hippocampus. After PHA-L injection into the caudal part of the medial septum or vertical limb of diagonal band of Broca, labeled terminals were observed predominantly in the ventral hippocampus.

Animals↗
PubMed1993

Input from the inferior colliculus to medial olivocochlear neurons in the rat: a double label study with PHA-L and cholera toxin.

The inferior colliculus provides a strong descending influence capable of modulating the excitability levels of olivocochlear neurons (Rajan, 1990). In an attempt to anatomically demonstrate this pathway in rats, an experimental paradigm was designed by which anterogradely transported Phaseolus vulgaris-leucoagglutinin (PHA-L), which delineates axonal arbors, and retrogradely transported cholera toxin B subunit alone (CT-B) or conjugated to horseradish peroxidase (CT-HRP), which delineate dendritic arbors, are visualized in the same brainstem sections. PHA-L was injected unilaterally into the central nucleus of the inferior colliculus of adult rats 5-9 days prior to injection of CT-B or CT-HRP into either the contralateral or the ipsilateral cochlea. Descending collicular axons labeled with PHA-L densely innervate the ventral nucleus of the trapezoid body (VNTB), which contains neurons of the medial olivocochlear system (MOCS), but do not enter the lateral superior olive, where the neurons of the lateral olivocochlear system (LOCS) are found. The collicular projection to VNTB is largely ipsilateral and supplies mostly the ventral half of the nucleus. Within VNTB, the collicular fibers intermingle with dendrites and, to a lesser extent, cell bodies of MOCS. Collicular boutons, predominantly of the en passant type, are often observed in close apposition to dendrites and, less frequently, cell bodies of both crossed and uncrossed MOCS. These light microscopic results suggest the existence of direct, synaptic contacts between descending collicular axons and ipsilateral crossed and uncrossed MOCS. Numerous collicular boutons were also seen at a distance from MOCS, suggesting that they establish synapses with other neuron types of the VNTB that do not send their axons to the cochlea.

Animals↗
PubMed1991

Chemiluminescence is increased in a subgroup of PHA-stimulated lymphocytes from patients with systemic lupus erythematosus and inhibited by 5-lipoxygenase inhibitors.

Chemiluminescence (CL) was examined in phytohemagglutinin (PHA)-stimulated control and lupus lymphocytes because oxidative radicals have the chemical potential to generate DNA changes recently observed in lupus lymphocytes. Increased CL was found in 30 of 65 PHA-stimulated lupus lymphocyte samples by a luminol assay. CL did not correspond statistically to oxidative potential measured by a nitroblue tetrazolium assay. CL did not appear to be related to disease activity, organ involvement, or drug therapy. However, six of six males tested had positive CL activity. Cocultivation of CL-positive PHA-stimulated lupus lymphocytes with metabolic inhibitors of various oxidative enzymes revealed that 50 microM arachidonic acid dramatically inhibited the excess oxidation. A specific inhibitor of 5-lipoxygenase activity, 3 microM nordihydroguaiaretic acid, abolished excess CL activity. These studies suggest that chemiluminescence assays can be used to better understand the oxidative metabolism in lupus lymphocytes. The enzyme 5-lipoxygenase may be dysfunctional in a subgroup of lupus patients.

Humans↗
PubMed2006

Polyhydroxyalkanoate (PHA) biosynthesis from structurally unrelated carbon sources by a newly characterized Bacillus spp.

A newly acquired polyhydroxyalkanoate (PHA) producing Bacillus spp. was identified to be a strain of Bacillus cereus using a range of microbiological and molecular techniques. This strain, named B. cereus SPV, was found to be capable of using a wide range of carbon sources including glucose, fructose, sucrose, various fatty acids and gluconate for the production of PHAs, an advantage for the commercial production of the polymers. The media used for the polymer production was novel in the context of the genus Bacillus. The PHA, once produced, was found to remain at a constant maximal concentration, without any degradation, a great advantage for the commercial production of the PHAs. This particular strain of Bacillus spp. was able to synthesize various PHAs with 3-hydroxybutyrate (3HB), 3-hydroxyvalerate (3HV) and 4-hydroxybutyrate (4HB)-like monomer units from structurally unrelated carbon sources such as fructose, sucrose and gluconate. This is the first report of the incorporation of a 4HB related monomer containing PHA by the genus Bacillus and from structurally unrelated carbon sources. The PHAs isolated had molecular weights ranging between (0.4 and 0.8) x 10(6) and low polydispersity index values (M(W)/M(N)) ranging from 2.6 to 3.4.

Bacillus cereus↗
PubMed1990

Efferent connections of the A1 noradrenergic cell group: a DBH immunohistochemical and PHA-L anterograde tracing study.

Immunohistochemical localization of the catecholamine biosynthetic enzymes tyrosine hydroxylase (TH), dopamine beta-hydroxylase (DBH), and phenylethanolamine N-methyltransferase (PNMT) was employed to reveal the anatomical organization of the A1 noradrenergic cell group in the caudal ventrolateral medulla oblongata of the rat. Subsequently, the supraspinal efferent axonal projections of A1 were investigated with a view to elucidating the anatomical substrates underlying its postulated function in central fluid and cardiovascular homeostasis. Within the caudal medulla, DBH-positive/PNMT-negative (noradrenergic) neurons were observed extending bilaterally through the ventrolateral medullary reticular formation from upper cervical spinal cord levels to the level of the area postrema. At the rostral pole of A1, its neurons intermingled with PNMT-immunoreactive perikarya of the more rostrally situated C1 adrenergic cell group. Discrete injections of the anterogradely transported plant lectin Phaseolus vulgaris leucoagglutinin (PHA-L) into A1 resulted in terminal labeling in a number of presumptive efferent target sites including the nucleus of the solitary tract, rostral ventrolateral medulla, dorsal parabrachial nucleus, Kolliker-Fuse nucleus, central grey, dorsomedial nucleus of the hypothalamus, perifornical region, zona incerta, lateral hypothalamus, paraventricular nucleus of the hypothalamus, supraoptic nucleus, bed nucleus of the stria terminalis, and organum vasculosum of the lamina terminalis. Tissue sections adjacent to those reacted for PHA-L were processed immunohistochemically for DBH to determine if anterogradely labeled terminals were localized in regions that demonstrated appropriate immunoreactivity. The majority of regions in which PHA-L terminal labeling was present also exhibited moderate to intense DBH activity. These experiments provide neuroanatomical evidence for direct efferent pathways from the A1 noradrenergic cell group to a number of supraspinal sites that have been reliably implicated in the neural circuitry underlying the central regulation of fluid and cardiovascular homeostasis. Furthermore, the results suggest a selective anatomical interrelation between A1 and sites in the basal forebrain and hypothalamus in which vasopressinergic neurons have been previously demonstrated. It is postulated that the noradrenergic A1 projections observed in this investigation represent the morphological substrate through which A1 exerts a significant influence on cardiovascular regulatory mechanisms.

Animals↗
PubMedDate not supplied

Microbial inclusions with special reference to PHA inclusions and intracellular boundary envelopes.

Polyhydroxyalkanoates (PHAs), in the form of metabolic storage reserves, are assembled in intracellular cytoplasmic inclusions, often called granules. This review discusses both the structure and function of this assembly. In addition an overview of other microbial cellular inclusions is presented. This is not a compilation of all such structures but a description of those that are similar in many ways to either the structure or function of the PHA inclusions and are made up of monolayer envelopes and their storage compounds. Not unique, such inclusions provide many similar examples which, in turn, provide useful analogies to the PHA inclusions. A study of the PHA inclusions has been carried out in a comparative electron microscope examination and by protein analysis of a number of organisms and E. coli transformants.

Bacteria↗
PubMedDate not supplied

PHA production, from bacteria to plants.

The genes encoding the polyhydroxyalkanoate (PHA) biosynthetic pathway in Ralstonia eutropha (3-ketothiolase, phaA or bktB; acetoacetyl-CoA reductase, phaB; and PHA synthase, phaC) were engineered for plant plastid targeting and expressed using leaf (e35S) or seed-specific (7s or lesquerella hydroxylase) promoters in Arabidopsis and Brassica. PHA yields in homozygous transformants were 12-13% of the dry mass in homozygous Arabidopsis plants and approximately 7% of the seed weight in seeds from heterozygous canola plants. When a threonine deaminase was expressed in addition to bktB, phaB and phaC, a copolyester of 3-hydroxybutyrate and 3-hydroxyvalerate was produced in both Arabidopsis and Brassica.

Acetyl-CoA C-Acyltransferase↗
PubMed2000

Development of an improved in vitro activity assay for medium chain length PHA polymerases based on CoenzymeA release measurements.

An improved activity assay for polyhydroxyalkanoate (PHA) polymerases from Pseudomonas oleovorans GPo1 was developed. The activity assay is based on the detection of released Coenzyme A (CoA) using 5,5'-dithiobis (2-nitrobenzoic acid) (DTNB), a compound which specifically reacts with thiol groups. The formed adduct was measured spectrophotometrically with high sensitivity and accuracy. The assay was used to study the effect of several additives on the activity of granule-associated PHA polymerase. Mild non-ionic detergents such as Tween-20, Triton X-100, CHAPS and Hecameg all appeared to be strongly inhibitory. In contrast, bovine serum albumin (BSA) had a strong stimulatory effect on the activity and stability of the PHA polymerases. Using optimized conditions, activities up to 5.8 U/mg granule-bound polymerase have been measured.

Acyltransferases↗
PubMed1997

Ouabain effects on activated lymphocytes: augmentation of CD25 expression on TPA-stimulated cells and of CD69 on PHA-and TPA-stimulated cells.

Ouabain (OUA) was capable of inhibiting peripheral blood lymphocyte (PBL) proliferation induced by phyothaemagglutinin (PHA) of phorbol ester (TPA), as measured by thymidine incorporation or cell cycle analysis. In this latter case it was possible to detect a block in the progression from G1 to S phase. This inhibition could not be reversed by interleukin (IL)-2 and was not due to an effect on CD 25 expression, as this molecule was only reduced in PHA cultures treated with OUA. Conversely, cultures activated by TPA and OUA showed an increased expression of CD25. The activation antigen CD69 was increased in both situation, suggesting that despite the absence of proliferative response the cells were being activated. The possibility that these cells were being deviated to the activation pathway leading to apoptosis is now under investigation. This study also suggested that CD25 induction may occur via different pathways, and that the selective effect of OUA for PHA-activated cells may become a useful tool for the understanding of the process.

Antigens, CD↗
PubMedDate not supplied

High and low density PHA- (but not ConA-) activated T cells stimulate the autologous mixed lymphocyte reaction.

In this study, PHA- and ConA-activated cells (PAC and CAC) were used as stimulators in mixed lymphocyte reactions (MLR) using autologous (auto) and allogeneic (allo) peripheral mononuclear cells as responders. PAC, but not CAC, were stimulatory in allo- and auto-MLR, and this stimulation was not due to residual PHA. In PAC which have been activated for 96 h, auto-MLR was due to determinants present on low density T-cell blasts, while with PAC which had been stimulated for more than 192 h, the determinants seemed to be associated with high density T cells. Anti-T3 monoclonal antibodies and certain anti-DR suppressed auto- and allo-MLR mediated by PAC when present throughout the entire MLR assays. CAC suppressed PAC-mediated auto-MLR in a dose-dependent fashion. This inhibition was not DR-restricted and was reversed by the addition of exogenous IL-2. Our results indicate that: depending upon the length of activation, both low density and high density PHA-activated T cells exhibited strong stimulatory capacity in auto-MLR; ConA-activated T cells failed to stimulate auto- or allo-MLR and suppressed MLR mediated by PAC; this suppression was due to suppressor cells, not to suppressor factors, and was readily reversed by exogenous IL-2; pretreatment of CAC with anti-TAC did not reverse the inhibition.

Antigens, Differentiation, T-Lymphocyte↗
PubMed2004

Snapper (Pagrus auratus) leucocyte proliferation is synergistically enhanced by simultaneous stimulation with LPS and PHA.

Important channels of communication between mammalian leucocytes have long been recognised. Here, data are reported that suggest similar integrations may occur between snapper leucocytes upon mitogen stimulation. Cell surface immunoglobulin (IgM) expression was used in conjunction with intracellular fluorescence staining and flow cytometry to differentiate proliferating peripheral blood leucocyte subsets (PBLs). Independent activation using phytohaemagglutinin (PHA) or lipopolysaccharide (LPS) drove both mIg(-)and mIg(+)cells into cycle. It is not known if the proliferation of mIg(+)cells was mediated by a mutually exclusive effect of the mitogen on each cell population, cognate cellular interaction or a soluble growth factor. Simultaneous activation of PBLs with PHA and LPS consistently induced significantly more cells to proliferate than the sum of proliferating cells stimulated solely with PHA or LPS. Together, the results suggest that different leucocyte subsets have the capability to influence their respective responses to mitogenic stimulation. Therefore, like in the mammalian immune system, communication may occur between snapper leucocyte subsets.

Animals↗
PubMed1973

Immunological studies in acute myeloid leukaemia: PHA responsiveness and serum inhibitory factors.

Sera from 16 of 20 patients with AML at some stage of the disease inhibited the in vitro PHA transformation of normal lymphocytes assessed by measuring the rate of DNA synthesis after 67-70 hours; 42% of pretreatment sera were inhibitory. Inhibitory activity was overcome at PHA concentrations 2-3 times greater than the concentration which allowed maximum discrimination between NHS and leukaemia sera.PHA transformation of washed lymphocytes obtained from AML patients before treatment and when receiving induction or consolidation (cytoreductive) chemotherapy was reduced only when cultures contained a high proportion of primitive cells. Even in primitive cell contaminated cultures significant responses to PHA could be measured if conditions were modified to prevent increasing acidity.Reports of reduced in vitro immunological reactions in pretreatment and poor prognosis patients may therefore be due to the presence of primitive cells in culture, and in treated patients to the failure of chemotherapy to reduce the circulating primitive cell count. Serum inhibitory factors may have a significant immunosuppressive effect in vivo, but the accurate assessment of the role of immune mechanisms in AML should attempt the measurement of specific immunity.

Culture Techniques↗
PubMed1978

Fluorescence-polarization changes in mononuclear blood leucocytes after PHA incubation: differences in cells from patients with and without neoplasia.

In 32 healthy blood donors, 20 patients with histologically verified cancer and 18 patients with non-neoplastic diseases, the fluorescence polarization changes of fluorescein samples incorporated in mononuclear leucotyes were measured after incubation with PHA. The leucocytes of healthy persons, and 16/18 persons with non-neoplastic diseases, responded with a decrease in the degree of fluorescence polarization by about 20% from that in non-PHA-stimulated cells. In 19/20 patients with a variety of malignant tumours, the leucotyes did not respond to PHA stimulation with such a decrease. The exceptions among the patients with neoplastic and non-neoplastic diseases are considered, and may not be "false-negative" or "false-positive" respectively, but indicative of a particular situation in that disease. The biophysical mechanisms underlying the observed changes remain to be investigated.

Adult↗
PubMed1971

Cellular recognition in vitro by mouse lymphocytes. Effects of neonatal thymectomy and thymus graft restoration on alloantigen and PHA stimulation of whole and gradient-separated subpopulations of spleen cells.

The effects of thymectomy and thymus graft restoration upon the in vitro primary responses to alloantigens and PHA have been studied. It has been found that neonatal thymectomy substantially eliminates both PHA reactivity and responsiveness to alloantigens assayed in vitro in host spleen cell populations. Analysis of albumin density gradient-separated subpopulations of the spleen and thymus in such animals was also performed. It was found that the total and proportional representation of the individual density subpopulations was identical in neonatally thymectomized, in normal, and in thymectomized and thymus graft-restored animals. Therefore, thymectomized mice appear to retain a nonfunctioning, small, dense, lymphocyte population. Reconstitution of thymic-dependent in vitro reactivity was nearly complete when syngeneic, but not allogeneic or semisyngeneic thymus was employed. Occasional partial restoration did occur when F(1) thymus was employed, but never when allogeneic thymus was grafted. The grafted thymus contained PHA and alloantigen-reactive cells in a large, less dense B layer subpopulation, whereas the restored animals, as in the case of normals, showed these reactivities to be a property of a small, more dense cell population.

Albumins↗
PubMed2001

PHA-induced cell proliferation rescues human peripheral blood lymphocytes from X-ray-induced apoptosis.

Human peripheral blood G(0) lymphocytes were X-irradiated and allowed to recover for different periods both in the presence and absence of phytohemagglutinin (PHA). For each experimental condition the induction of apoptosis was investigated by nuclear morphology and formation of both DNA laddering and high molecular weight DNA fragments by pulsed field gel electrophoresis. The results showed that PHA cell growth stimulation could rescue peripheral blood lymphocytes (PBLs) from X-ray-induced apoptotic cell death. Instead, most X-irradiated lymphocytes held in G(0) phase, once they were committed to apoptosis, inexorably executed the process. These data indicate that the proliferative status of PBLs can influence apoptotic cell death: PHA-stimulated PBLs appear to be more radioresistant as a result of a less efficient apoptotic process. Therefore, in current tests for mutagenicity or cytotoxicity and in biodosimetrical studies the possible role of apoptosis has to be considered.

Apoptosis↗