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PubMed1994

Projections of the dorsal raphe nucleus to the brainstem: PHA-L analysis in the rat.

Early studies that used older tracing techniques reported exceedingly few projections from the dorsal raphe nucleus (DR) to the brainstem. The present report examined DR projections to the brainstem by use of the anterograde anatomical tracer Phaseolus vulgaris leucoagglutinin (PHA-L). DR fibers were found to terminate relatively substantially in several structures of the midbrain, pons, and medulla. The following pontine and midbrain nuclei receive moderate to dense projections from the DR: pontomesencephalic central gray, mesencephalic reticular formation, pedunculopontine tegmental nucleus, medial and lateral parabrachial nuclei, nucleus pontis oralis, nucleus pontis caudalis, locus coeruleus, laterodorsal tegmental nucleus, and raphe nuclei, including the central linear nucleus, median raphe nucleus, and raphe pontis. The following nuclei of the medulla receive moderately dense projections from the DR: nucleus gigantocellularis, nucleus raphe magnus, nucleus raphe obscurus, facial nucleus, nucleus gigantocellularis-pars alpha, and the rostral ventrolateral medullary area. DR fibers project lightly to nucleus cuneiformis, nucleus prepositus hypoglossi, nucleus paragigantocellularis, nucleus reticularis ventralis, and hypoglossal nucleus. Some differences were observed in projections from rostral and caudal parts of the DR. The major difference was that fibers from the rostral DR distribute more widely and heavily than do those from the caudal DR to structures of the medulla, including raphe magnus and obscurus, nucleus gigantocellularis-pars alpha, nucleus paragigantocellularis, facial nucleus, and the rostral ventrolateral medullary area. A role for the dorsal raphe nucleus in several brainstem controlled functions is discussed, including REM sleep and its events, nociception, and sensory motor control.

Animals↗
PubMed1994

Organization of the efferent projections from the pontine parabrachial area to the bed nucleus of the stria terminalis and neighboring regions: a PHA-L study in the rat.

The organization of efferent projections from the pontine parabrachial (pPB) area to the forebrain rostral to the central nucleus of the amygdala (Ce) was studied in the rat by using microinjections of Phaseolus vulgaris leucoagglutinin (PHA-L), into subregions of the pPB area. The present study is a follow-up of a former study (Bernard et al. [1993] J. Comp. Neurol. 329:201-229) which examines pPB projections onto the Ce. The results demonstrate that: (1) the pPB(m) region (the medial, the ventral lateral subnuclei and the waist area) diffusely projects to the lateral division (BSTL) of the bed nucleus of the stria terminalis (BST), the Ce-BSTL continuum (including, the dorsal portion of substantia innominata, the ventral portion of globus pallidus, the fundus striatum, and the substriatal area) and to a lesser extent the agranular insular cortex; (2) the pPB(1) region [the central lateral (pPBcl) and the outer portion of external lateral subnuclei] densely projects to the dorsal lateral subnucleus of BST (BSTdl); only the pPBcl subnucleus projects to the median, the anteroventral and the periventricular nuclei of the preoptic hypothalamus; and (3) the remaining pPB area (the dorsal lateral, part of the external lateral and the external medial subnuclei) projects to the nucleus of horizontal limb of diagonal band but does not project onto the BST and the preoptic hypothalamus. It is suggested that the pPB(m)-BSTL "diffuse pathway" is mainly implicated in motivational and autonomic aspects of taste. The pPB(1)-BSTdl and hypothalamic "concentrated pathways" could be implicated in autonomic and nociceptive processes.

Amygdala↗
PubMed1995

Organization of the efferent projections from the spinal cervical enlargement to the parabrachial area and periaqueductal gray: a PHA-L study in the rat.

The organization of efferent projections from the spinal cervical enlargement to the parabrachial (PB) area and the periaqueductal gray (PAG) was studied in the rat by using microinjections of Phaseolus vulgaris-leucoagglutinin (PHA-L) into different laminae around the C7 level. The results demonstrated two areas of cervical enlargement which project in different ways to the PB area and PAG. First, the superficial laminae (I, II) showed a very dense projection, with a clear contralateral dominance at the coronal level where the inferior colliculus merges with the pons, to a restricted "superficial" portion of the PB area, namely the lateral crescent area, the dorsal lateral, the superior lateral (PBsl), and the outer portion of the external lateral PB subnuclei. Less dense projections were observed in the Kölliker-Fuse nucleus (KF) and in the ventrolateral/lateral quadrant of the caudal and mid PAG. By contrast, the labeling was weak or absent in the other PB subnuclei and the outer adjacent regions; in particular, no, or very little, labeling was found in the cuneiform nucleus. The PB area appeared to be the supraspinal target that received the densest projection from laminae I and II. Projections were less dense in the PAG and the thalamus and markedly less in other sites such as the ventrolateral medulla, the subnucleus reticularis dorsalis, and the nucleus of the solitary tract. Second, the reticular portion of lamina V, the medial portion of laminae IV-VI up to X and lamina VIII, showed bilateral projections with a weak ipsilateral dominance and a high to medium density on a very restricted portion of the PB area, namely the internal lateral PB subnucleus. A lesser projection was also observed in the adjacent portion of the PBsl, the KF, and the lateral quadrant of the PAG. These results suggest that signals carried by neurons from lamina I-II converge on a restricted superficial portion of the PB area and the ventral part of the lateral quadrant of the PAG. These results are discussed in the context of the role of the spino-PB and spino-PAG pathways in nociception.

Animals↗
PubMed1995

Ascending projections of the posterior nucleus of the hypothalamus: PHA-L analysis in the rat.

With the exception of a report by R.B. Veazey, D.G. Amaral, and W.M. Cowan (1982, J. Comp. Neurol. 207:135-156) that examined the projections of the posterior hypothalamic area in the monkey by using the autoradiographic technique, the ascending projections of the posterior nucleus (PH) of the hypothalamus have not been systematically examined in any species. The present report describes the ascending projections of PH in the rat by using the anterograde anatomical tracer, Phaseolus vulgaris-leucoagglutinin (PHA-L). The major ascending route for PH fibers is the medial forebrain bundle. PH fibers project densely to several subcortical and cortical sites. The subcortical sites are the subthalamus/hypothalamus (zona incerta, the supramammillary nucleus, lateral, perifornical, dorsal, and anterior nuclei/areas), the thalamus (lateroposterior, laterodorsal, parafascicular, reuniens, paraventricular, central medial, paracentral, central lateral and intermediodorsal nuclei), the amygdala (central, lateral, and medial nuclei), the septal area (bed nucleus of stria terminalis, medial and lateral septum), and the basal forebrain (horizontal/vertical limbs of diagonal band nuclei and lateral preoptic area). The cortical sites are the perirhinal, insular, frontal (lateral agranular), prelimbic, and infralimbic cortices. The diversity of PH projections to subcortical and cortical "limbic-related" sites and to several structures with direct input to the hippocampus (supramammillary nucleus, reuniens, paraventricular and laterodorsal nuclei of the thalamus, medial and lateral septum, and perirhinal cortex) suggest that the PH may serve a critical role in various components of emotional behavior, including mnemonic processes associated with significant emotional events.

Amygdala↗
PubMed2003

Projections from the periamygdaloid cortex to the amygdaloid complex, the hippocampal formation, and the parahippocampal region: a PHA-L study in the rat.

The periamygdaloid cortex, an amygdaloid region that processes olfactory information, projects to the hippocampal formation and parahippocampal region. To elucidate the topographic details of these projections, pathways were anterogradely traced using Phaseolus vulgaris leukoagglutinin (PHA-L) in 14 rats. First, we investigated the intradivisional, interdivisional, and intra-amygdaloid connections of various subfields [periamygdaloid subfield (PAC), medial subfield (PACm), sulcal subfield (PACs)] of the periamygdaloid cortex. Thereafter, we focused on projections to the hippocampal formation (dentate gyrus, hippocampus proper, subiculum) and to the parahippocampal region (presubiculum, parasubiculum, entorhinal, and perirhinal and postrhinal cortices). The PACm had the heaviest intradivisional projections and it also originated light interdivisional projections to other periamygdaloid subfields. Projections from the other subfields converged in the PACs. All subfields provided substantial intra-amygdaloid projections to the medial and posterior cortical nuclei. In addition, the PAC subfield projected to the ventrolateral and medial divisions of the lateral nucleus. The heaviest periamygdalohippocampal projections originated in the PACm and PACs, which projected moderately to the temporal end of the stratum lacunosum moleculare of the CA1 subfield and to the molecular layer of the ventral subiculum. The PACm also projected moderately to the temporal CA3 subfield. The heaviest projections to the entorhinal cortex originated in the PACs and terminated in the amygdalo-entorhinal, ventral intermediate, and medial subfields. Area 35 of the perirhinal cortex was lightly innervated by the PAC subfield. Thus, these connections might allow for olfactory information entering the amygdala to become associated with signals from other sensory modalities that enter the amygdala via other nuclei. Further, the periamygdalohippocampal pathways might form one route by which the amygdala modulates memory formation and retrieval in the medial temporal lobe memory system. These pathways can also facilitate the spread of seizure activity from the amygdala to the hippocampal and parahippocampal regions in temporal lobe epilepsy.

Amygdala↗
PubMed1986

Abnormalities of T cells isolated from mediastinal lymph nodes and peripheral blood of patients with lung carcinoma: deficit in PHA-induced expression of HLA class II antigens and in autologous mixed lymphocyte reactions.

A reduced percentage of T cells isolated from mediastinal lymph nodes and peripheral blood of patients with lung carcinoma acquired HLA Class II antigens following in vitro stimulation with PHA. Furthermore T cells were functionally abnormal in autologous and allogeneic mixed lymphocyte reactions (MLR). The immunoregulatory properties of HLA Class II antigens and autologous MLRs suggest that these abnormalities may affect the interaction of the host's immune system with tumor cells.

Adult↗
PubMed1982

Genetic analysis of human lymphocyte proteins by two-dimensional gel electrophoresis: 3. Frequent occurrence of genetic variants in some abundant polypeptides of PHA-stimulated peripheral blood lymphocytes.

The 100 or so most intensely Coomassie blue-stained polypeptides from PHA-stimulated peripheral blood lymphocytes were analyzed by two-dimensional electrophoresis in combination with family and population studies. Besides polymorphic lymphocyte cytosol 64k polypeptide reported previously, genetic variants were frequently observed in three polypeptides with molecular weights of 100,000, 49,000, and 40,000. All of them occur in the cytosol. These variant polypeptides are charge variants, because they are separated in the isoelectric focusing dimension. It is indicated by family and population studies and cell distribution analysis that the polypeptide with a molecular weight of 100,000 shows a genetic polymorphism determined by two alleles at a new autosomal locus, as described in the following paper. Family and population studies also suggest that a genetic polymorphism defined by alleles at an autosomal locus is present in each of the polypeptides with molecular weights of 49,000 and 40,000. In contrast to the previous reports of the extremely restricted genetic variability of the 100 or so most abundant fibroblast polypeptides, the present data indicate that common genetic variants are present at least in four of the 100 or so most intensely Coomassie blue-stained lymphocyte polypeptides. The result also shows that careful side-by-side comparison of two-dimensional electrophoresis patterns among both parents and their children is an effective method to detect genetic variant polypeptides.

Blood Protein Electrophoresis↗
PubMed1976

The human leukocyte test system. IX. DNA synthesis and mitoses in PHA-stimulated 2-day cultures set up with Eagle's mimimal essential medium (MEM).

In human leukocyte cultures set up with Eagle's MEM and stimulated with Difco's PHA M, DNA synthesis and mitotic indices were analyzed by means of 3(H)-thymidine autoradiography and cell counting from 23 up to 52 h after culture initiation. Considerable amounts of DNA synthesis and mitoses were found in this time span. This resembles the results found with Ham's F-10 medium. However, the DNA synthesis pattern and the distribution of mitotic indices a higher yield of asynchrony in Eagle's MEM as compared with Ham's F-10 cultures. Proportions of first, second, and third mitoses at 72 h culture time were determined with different methods.

Culture Media↗
PubMed1983

Suppression of PHA-induced human T-lymphocyte colony formation following in vivo administration of anti-lymphocyte globulin. Report of one case.

The administration of anti-lymphocyte globulin in a patient with pure red cell aplasia completely abrogated the PHA-induced T colony forming capacity of peripheral blood mononuclear cells. The possible mechanisms of the ALG-induced suppression are discussed. It is proposed that T-colony assays may represent an useful tool for monitoring some effects of immunosuppressive drugs on T cells.

Anemia, Aplastic↗
PubMed1980

New complex Ph' translocation t (10; 14; 22) in bone marrow cells and in PHA-stimulated peripheral blood cultures in chronic myelocytic leukaemia.

A patient with chronic myelocytic leukaemia (CML) and a new complex Philadelphia chromosome (Ph') translocation, t (10; 14; 22), is described. This three way Ph' translocation not involving chromosome 9 was present in the majority of the bone marrow cells, as well as in a great proportion of metaphases from phytohaemagglutinin (PHA) stimulated peripheral blood cultures. The possibility that the Ph' translocation was present also in lymphocytes is discussed and at this regard the involvement of chromosome 14 is of interest considering the documented non random involvement of chromosome 14 in lymphoid malignancies.

Aged↗
PubMed1980

The use of a passive hemagglutination test (PHA) in the diagnosis of viral eye diseases. Investigation of lacrimal fluid for the presence of antibody to herpes simplex virus (HSV).

The use of a passive haemagglutination test (PHA) with a stable erythrocyte diagnostic preparation sensitized with herpes simplex virus for the detection of antibody to herpes simplex virus (HSV) in the lacrimal fluid of the patients is described. Specific antibody to HSV antigen was found in 68% of the lacrimal fluids taken from patients suffering from herpetic eye disease. In patients with superficial keratitis the antibodies were found more frequently and in higher titers than in deep involvements of the cornea. In patients with keratitis and keratouveitis of unknown etiology, antibody in tears was found in 23%, and in those with inflammatory diseases of nonherpetic etiology, in 14.2%. In clinically normal subjects and patients with non-inflammatory eye affections no HSV antibody could be found in tears. Geometric mean antibody titers in tears of patients with herpes simplex keratitis and keratouveitis were statistically significantly higher than in patients with other diagnoses. The diagnostic importance of HSV antibody found in tears is discussed.

Adolescent↗
PubMed1987

Role of monocytes in the inhibitory effect of calcitriol on PHA-stimulated lymphocytes.

The possible role played by monocytes in the inhibitory effect of calcitriol on phytohemagglutinin (PHA)-stimulated lymphocyte proliferation was assessed by testing the effect of this sterol under different cell culture conditions. Calcitriol had a dose-dependent inhibitory effect on lymphocyte proliferation in concentrations ranging from 10(-10) up to 10(-8) M. The effect of 10(-9) M calcitriol was almost completely abolished by: a) monocyte depletion, b) inhibition of prostaglandin (PG) synthesis by indomethacin, and c) addition of exogenous interleukin-2 (IL-2). These results suggested that the inhibitory effect of calcitriol was mediated through monocytes. This possibility was substantiated by the following observations: a) the calcitriol inhibitory effect was restored when autologous adherent cells were added to monocyte-depleted PBM cells; b) the supernatant of adherent cells cultured for 24 hours in the presence of calcitriol exerted a marked inhibitory effect on lymphocyte proliferation; and c) this effect was not longer evident when adherent cells were cultured in the presence of calcitriol plus indomethacin. These data support the hypothesis that calcitriol acts, at least partially, through the monocytes, inducing an increased release of PG, with subsequent inhibition of IL-2 synthesis, then resulting in a decreased lymphocyte proliferation.

Calcitriol↗
PubMed1996

Expression of L-PHA-binding proteins in breast cancer: reconstitution and molecular characterization of beta 1-6 branched oligosaccharides in three-dimensional cell culture.

Expression of beta 1-6 branched oligosaccharides in human breast cancer cells was investigated in vivo and in vitro. Lectin histochemical and lectin blotting analyses of surgically resected specimens were performed using L-PHA (phaseolus vulgaris leukoagglutinin) lectin, which binds to beta 1-6 oligosaccharides. The glycoproteins bearing beta 1-6 oligosaccharides of breast cancer tissues were found to be 170 kD and 120 kD in molecular weight, and the former appeared to be an epitope of carcinoembryonic antigen (CEA). The beta 1-6 oligosaccharides were expressed in both cancer cell lines at the outer layer of the colonies when cultured in type I collagen, but not in agarose gel. No correlation was observed between beta 1-6 expression and cell cycle. The beta 1-6 oligosaccharides did not coincide with breast cancer-associated antigens, such as CEA, MUC1, and cathepsin D. The beta 1-6 oligosaccharides of these cell lines were markedly inhibited when swainsonine, a mannosidase II inhibitor, was added to the culture medium. The 120 kD molecule, which was obtained from MCF-7 cells cultured in type I collagen gel, was consistent with that of breast cancer tissues and was similar to lysosome-associated membrane glycoproteins (LAMPs). The results suggest that the glycoproteins bearing beta 1-6 branched oligosaccharides in human breast cancer incorporate an epitope of CEA and human LAMPs and that the expression of LAMPs may depend on their surrounding matrices and may play an important role in cancer invasion or metastasis.

Breast Neoplasms↗
PubMed1979

Inhibitory effect of cytokinins on PHA-induced human lymphocyte stimulation.

The inhibitory effect of various purine derivatives on PHA-induced human lymphocyte blast formation was studied. Two nucleoside cytokinins, N6-benzyladenosine and N6-isopentenyladenosine, inhibited blast formation at concentrations as low as 10(-6) M. However, the other cytokinins, which lacked the ribosyl residue at N9 position, had to be at the higher molar concentration of 10(-4) before they could induce the same inhibitory effect.

Adenosine↗
PubMed1984

An in vitro effect of an acute non-specific inflammatory serum on rat lymphocytes proliferative response to PHA.

This study has demonstrated that serum obtained from animals undergoing an acute inflammatory reaction induced by an intrapleural injection of dextran is able to modulate the proliferative response to PHA of lymph node and spleen cells in vitro. This response is dependent on the concentration of the inflammatory serum and on the time of collection of the serum during the acute inflammatory process. At low concentrations of serum (0.5%) stimulatory activity was observed at all time points. At higher concentrations (1%) inhibitory activity was present in 24 and 72 h sera. These results support the previous observation that the acute non-specific inflammatory reaction is able to modify the function of different cell types. It has been suggested that both stimulatory and inhibitory factors are present and the balance between these changes during the course of the inflammatory reaction.

Animals↗
PubMed1986

Action of histamine on PHA chemiluminescence response of blood mononuclear cells in autoimmune patients.

Peripheral blood mononuclear cells (PBMs) of patients suffering from autoimmune diseases showed significantly lower chemiluminescence response to PHA than the controls. High doses of histamine (10(-5) M) inhibited the chemiluminescence while low doses caused a mild enhancement in all groups of patients. The preformed histamine content of the PBMs was not significantly higher in the patients than in the controls. Individually, a reverse correlation was found between the activity of the disease and the sensitivity of the cells to histamine. In very active cases, virtually no inhibition by histamine was found.

Arthritis, Rheumatoid↗