Search

The CFTR High Expresser (CHE) cells express eightfold higher levels of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl−channel compared with neighboring enterocytes and were first identified by our laboratory (Ameen et al., Gastroenterology 108: 1016, 1995). We used double-label immunofluorescence microscopy to further study these enigmatic epithelial cells in rat intestine in vivo or ex vivo. CHE cells were found in duodenum, most frequent in proximal jejunum, and absent in ileum and colon. CFTR abundance increased in CHE cells along the crypt-villus axis. The basolateral Na+K+Cl−cotransporter NKCC1, a key transporter involved in Cl−secretion, was detected at similar levels in CHE cells and neighboring enterocytes at steady state. Microvilli appeared shorter in CHE cells, with low levels of Myosin 1a, a villus enterocyte-specific motor that retains sucrase/isomaltase in the brush-border membrane (BBM). CHE cells lacked alkaline phosphatase and absorptive villus enterocyte BBM proteins, including Na+H+exchanger NHE3, Cl−/HCO3−exchanger SLC26A6 (putative anion exchanger 1), and sucrase/isomaltase. High levels of the vacuolar-ATPase proton pump were observed in the apical domain of CHE cells. Levels of the NHE regulatory factor NHERF1, Na-K-ATPase, and Syntaxin 3 were similar to that of neighboring enterocytes. cAMP or acetylcholine stimulation robustly increased apical CFTR and basolateral NKCC1 disproportionately in CHE cells relative to neighboring enterocytes. These data strongly argue for a specialized role of CHE cells in Cl−-mediated “high-volume” fluid secretion on the villi of the proximal small intestine.

Changes in intestinal luminal pH affect mucosal ion transport. The aim of this study was to compare how luminal pH and specific second messengers modulate the membrane traffic of four major ion transporters (CFTR, NHE3, NKCC1, and NBCe1) in rat small intestine. Ligated duodenal, jejunal, and ileal segments were infused with acidic or alkaline saline, 8-Br-cAMP, or the calcium agonist carbachol in vivo for 20 min. Compared with untreated intestine, lumen pH was reduced after cAMP or carbachol and increased following HCO3−-saline. Following HCl-saline, lumen pH was restored to control pH levels. All four secretory stimuli resulted in brush-border membrane (BBM) recruitment of CFTR in crypts and villi. In villus enterocytes, CFTR recruitment was coincident with internalization of BBM NHE3 and basolateral membrane recruitment of the bicarbonate transporter NBCe1. Both cAMP and carbachol recruited NKCC1 to the basolateral membrane of enterocytes, while luminal acid or HCO3−retained NKCC1 in intracellular vesicles. Luminal acid resulted in robust recruitment of CFTR and NBCe1 to their respective enterocyte membrane domains in the upper third of the villi; luminal HCO3−induced similar membrane changes lower in the villi. These findings indicate that each stimulus promotes a specific transporter trafficking response along the crypt-villus axis. This is the first demonstration that physiologically relevant secretory stimuli exert their actions in villus enterocytes by membrane recruitment of CFTR and NBCe1 in tandem with NHE3 internalization.

We examined the cell-specific subcellular expression patterns for sodium- and potassium-coupled chloride (NaK2Cl) cotransporter 1 (NKCC1), Na+bicarbonate cotransporter (NBCe1), cystic fibrosis transmembrane conductance regulator (CFTR), and Na+/H+exchanger 3 (NHE3) to understand the functional plasticity and synchronization of ion transport functions along the crypt-villus axis and its relevance to intestinal disease. In the unstimulated intestine, all small intestinal villus enterocytes coexpressed apical CFTR and NHE3, basolateral NBCe1, and mostly intracellular NKCC1. All (crypt and villus) goblet cells strongly expressed basolateral NKCC1 (at approximately three-fold higher levels than villus enterocytes), but no CFTR, NBCe1, or NHE3. Lower crypt cells coexpressed apical CFTR and basolateral NKCC1, but no NHE3 or NBCe1 (except NBCe1-expressing proximal colonic crypts). CFTR, NBCe1, and NKCC1 colocalized with markers of early and recycling endosomes, implicating endocytic recycling in cell-specific anion transport. Brunner's glands of the proximal duodenum coexpressed high levels of apical/subapical CFTR and basolateral NKCC1, but very low levels of NBCe1, consistent with secretion of Cl−-enriched fluid into the crypt. The cholinergic agonist carbachol rapidly (within 10 min) reduced cell volume along the entire crypt/villus axis and promoted NHE3 internalization into early endosomes. In contrast, carbachol induced membrane recruitment of NKCC1 and CFTR in all crypt and villus enterocytes, NKCC1 in all goblet cells, and NBCe1 in all villus enterocytes. These observations support regulated vesicle traffic in Cl−secretion by goblet cells and Cl−and HCO3−secretion by villus enterocytes during the transient phase of cholinergic stimulation. Overall, the carbachol-induced membrane trafficking profile of the four ion transporters supports functional plasticity of the small intestinal villus epithelium that enables it to conduct both absorptive and secretory functions.

The actin motor myosin VI regulates endocytosis of cystic fibrosis transmembrane conductance regulator (CFTR) in the intestine, but the endocytic adaptor linking CFTR to myosin VI is unknown. Dab2 (Disabled 2) is the binding partner for myosin VI, clathrin, and alpha-AP-2 and directs endocytosis of low density lipoprotein receptor family members by recognizing a phosphotyrosine-binding domain. However, CFTR does not possess a phosphotyrosine-binding domain. We examined whether alpha-AP-2 and/or Dab2 were binding partners for CFTR and the role of myosin VI in localizing endocytic adaptors in the intestine. CFTR co-localized with alpha-AP-2, Dab2, and myosin VI and was identified in a complex with all three endocytic proteins in the intestine. Apical CFTR was increased in the intestines of Dab-2 KO mice, suggesting its involvement in regulating surface CFTR. Glutathione S-transferase pulldown assays revealed binding of CFTR to alpha-AP-2 (but not Dab2) in the intestine, whereas Dab-2 interacted with alpha-AP-2. siRNA silencing of alpha-AP-2 in cells significantly reduced CFTR endocytosis, further supporting alpha-AP-2 as the direct binding partner for CFTR. alpha-AP-2 and Dab2 localized to the terminal web regions of enterocytes, but Dab2 accumulated in this location in Snell's Waltzer myosin VI((sv/sv)) intestine. Ultrastructural examination revealed that the accumulation of Dab2 correlated with prominent involution and the loss of normal positioning of the intermicrovillar membranes that resulted in expansion of the terminal web region in myosin VI((sv/sv)) enterocytes. The findings support alpha-AP-2 in directing myosin VI-dependent endocytosis of CFTR and a requirement for myosin VI in membrane invagination and coated pit formation in enterocytes.

We previously demonstrated greater HPA axis activation in adult men compared to adult women following low-dose administration of the anticholinesterase inhibitor, physostigmine (PHYSO). Because blood sampling was done infrequently following PHYSO, the rise times of AVP, ACTH1-39, and cortisol could not be determined. In the present study, we determined the sequence of hormone increases by frequent blood sampling following PHYSO. Twelve adult women and 12 adult men underwent three test sessions 5-7 days apart: PHYSO, saline control, and repeat PHYSO. As in the earlier study, PHYSO produced no side effects in half the subjects and mild side effects in the other half, with no significant female-male differences. None of the hormone responses was significantly correlated with the presence or absence of side effects. In both women and men, the AVP increase preceded the ACTH1-39 increase, which in turn preceded the cortisol increase. The AVP and ACTH AUCs were significantly positively correlated in both women and men, supporting AVP as an acute stimulus to ACTH secretion. Also as in the earlier study, the AVP response to PHYSO was more than twice as great in men as in women, but the difference was not statistically significant. We therefore analyzed the results of both studies combined (N=26 women and 26 men). The men had a significantly greater AVP response and a trend toward a greater ACTH1-39 response compared to the women. These findings further support the concept of sexual diergism (functional sex difference) in the influence of CNS cholinergic systems on HPA hormone secretion.

Multiple doses of the dietary supplement l -ephedrine can cause severe hyperthermia and modest dopamine depletions in the rat brain. Since d -amphetamine treatment can result in neurodegeneration, the potential of l -ephedrine to produce similar types of degeneration was investigated. Adult male rats, some implanted in the caudate/putamen (CPu) for microdialysis, were given four doses of 25 mg/kg l -ephedrine or 5 mg/kg d -amphetamine (2 h between doses) at an ambient temperature of 23 °C. l -ephedrine-induced degeneration in the forebrain was dependent on the degree of hyperthermia. Layer IV of the parietal cortex was the most sensitive to l -ephedrine treatment with peak body temperatures of at most 40.0 °C necessary to produce degeneration. Extensive neurodegeneration in the parietal cortex after l -ephedrine treatment was as pronounced as that previously described for d -amphetamine treatment and also occurred in the intralaminar, ventromedial and ventrolateral thalamic nuclei in rats with severe hyperthermia (peak body temperatures>41.0 °C). The neurodegeneration induced by l -ephedrine may have resulted in part from excitotoxic mechanisms involving the indirect pathways of the basal ganglia and related areas. No differences were observed between microdialysis and non-implanted rats with respect to degree of tyrosine hydroxylase (TH) loss in the CPu after either d -amphetamine or l -ephedrine treatment. However, neurodegeneration resulting from d -amphetamine and l -ephedrine was reduced in the microdialysis animals in the hemisphere ipsilateral to the probe, which raises concerns when using the technique of in vivo microdialysis to evaluate neurodegeneration. The results of this study, in conjunction with human clinical evaluation of ephedrine neurotoxicity, indicate that regionally specific damage may occur in the cortex of some humans exposed to ephedrine in the absence of stroke or hemorrhage.

Reorganization of the cerebellar glomerulus, the main synaptic complex within the granule cell layer, was investigated using quantitative morphological techniques. All afferents to the cerebellar cortex, including mossy-fibers, were surgically destroyed by undercutting the cerebellar vermis. Fifteen days after the operation, which resulted in the removal of the main excitatory afferent to the glomerulus, a significant reorganization of the whole synaptic complex was observed, whereas the structural integrity of the glomerulus was remarkably well preserved. This was indicated by the observation that the number of granule cell dendrites (approximately 50 per glomerulus), as well as the number of dendritic digits (approximately 210 per glomerulus) bearing most of the approximately 230 synaptic junctions per glomerulus, did not change significantly after mossy-fiber degeneration. The total number of synapses in the reorganized glomerulus did not change either, despite the disappearance of two-thirds of (excitatory) synaptic junctions caused by mossy-fiber degeneration. In the reorganized glomeruli, however, the inhibitory, GABA-containing Golgi axonal varicosities became the dominant synaptic type-about four-fifths (approximately 200) of all synapses within the glomerulus-whereas the dendritic synapses between the granule cells represented only one-fifth of all synaptic junctions. The quantitative data of the reorganized cerebellar glomerulus demonstrate both a remarkable constancy and a plasticity of the excitatory granule cells and inhibitory Golgi neurons building up this synaptic complex. Constancy (the preservation of certain specific structural features) is represented by an eventually unchanged number of dendrites and synaptic junctions within the deafferented glomerulus. Such constancy was made possible, however, by the morphogenetic plasticity of both nerve-cell types to produce new, dendro-dendritic and axo-dendritic synapses to compensate for the loss of mossy-fiber synapses.

The Brunner's glands of the proximal duodenum exert barrier functions through secretion of glycoproteins and antimicrobial peptides. However, ion transporter localization, function, and regulation in the glands are less clear. Mapping the subcellular distribution of transporters is an important step toward elucidating trafficking mechanisms of fluid transport in the gland. The present study examined 1) changes in the distribution of intestinal anion transporters and the aquaporin 5 (AQP5) water channel in rat Brunner's glands following second messenger activation and 2) anion transporter distribution in Brunner's glands from healthy and disease-affected human tissues. Cystic fibrosis transmembrane conductance regulator (CFTR), AQP5, sodium-potassium-coupled chloride cotransporter 1 (NKCC1), sodium-bicarbonate cotransporter (NBCe1), and the proton pump vacuolar ATPase (V-ATPase) were localized to distinct membrane domains and in endosomes at steady state. Carbachol and cAMP redistributed CFTR to the apical membrane. cAMP-dependent recruitment of CFTR to the apical membrane was accompanied by recruitment of AQP5 that was reversed by a PKA inhibitor. cAMP also induced apical trafficking of V-ATPase and redistribution of NKCC1 and NBCe1 to the basolateral membranes. The steady-state distribution of AQP5, CFTR, NBCe1, NKCC1, and V-ATPase in human Brunner's glands from healthy controls, cystic fibrosis, and celiac disease resembled that of rat; however, the distribution profiles were markedly attenuated in the disease-affected duodenum. These data support functional transport of chloride, bicarbonate, water, and protons by second messenger-regulated traffic in mammalian Brunner's glands under physiological and pathophysiological conditions.

The aromatase enzyme (estrogen synthetase) catalyzes the conversion of testosterone to estrogen in peripheral and central nervous tissue. Light and electron microscopic immunocytochemistry was used to study the localization of this enzyme in the septal area of adult male and female albino rats. Aromatase-immunoreactivity was found restricted to neuronal somata and dendritic arbors, and no sex differences were detected in its distribution or intensity. Most aromatase-immunoreactive neurons formed two oblique bands in the lateral and the medial zones of the lateral septum; in addition, labeled cells were present in the septohippocampal nucleus and the laterodorsal portion of the bed nucleus of the stria terminalis. Electron microscopy revealed that the majority of aromatase-positive neurons in the lateral septum exhibit somatic spines, a characteristic marker of a neuron population that is known to contribute to local and extraseptal projections. The presence of aromatase in lateral septal somatospiny neurons suggests that estrogen formed by these neurons may be critically involved in the septal control of steroid-dependent behaviors.

The aromatase (estrogen synthetase) enzyme catalyzes the conversion of androgens to estrogens in peripheral tissues, as well as in the brain. Our study aimed at comparing the brain distribution of aromatase-immunoreactive neurons in male and female, normal and gonadectomized rats. Light microscopic immunostaining was employed using a purified polyclonal antiserum raised against human placental aromatase. Two anatomically separate aromatase-immunoreactive neuronal systems were detected in the rat brain: A “limbic telencephalic” aromatase system was composed by a large population of labeled neurons in the lateral septal area, and by a continuous “ring” of neurons of the laterodorsal division of the bed nucleus of stria terminalis, central amygdaloid nucleus, stria terminalis, and the substantia inominata-ventral pallidum-fundus striati region. The other, “hypothalamic” aromatase system consisted of neurons scattered in a dorsolateral hypothalamic area including the paraventricular, lateral and dorsomedial hypothalamic nuclei, the subincertal nucleus as well as the zona incerta. In addition, a few axon-like processes (unresponsive to gonadectomy) were present in the preoptic-anterior hypothalamic complex, the ventral striatum, and midline thalamic regions. No sexual dimorphism was observed in the distribution or intensity of aromatase-immunostaining. However, 3 days, 2, 3, 8, 16, or 32 weeks after gonadectomy, aromatase-immunoreactive neurons disappeared from the hypothalamus, whereas they were still present in the limbic areas of both sexes. The results indicate the existence of two distinct estrogen-producing neuron systems in the rat brain: (1) a “limbic ring” of aromatase-labeled neurons of the lateral septum-bed nucleus-amygdala complex unresponsive to gonadectomy; and (2) a sex hormone-sensitive “hypothalamic” aromatase neuron system.

Electron microscopic immunocytochemistry was combined with acute anterograde axon degeneration, following transection of the fimbria-fornix, to describe the innervation of somatospiny neurons by vasopressin-immunoreactive and degenerated hippocamposeptal axon terminals in the rat lateral septal area. Vasopressin-immunopositive boutons characterized by symmetric synaptic membrane specializations, and the degenerated hippocamposeptal axon terminals which form asymmetric synaptic contacts, frequently terminate on the same dendritic and somatic profiles, and particularly on the somata of somatospiny neurons. Although hippocamposeptal fibers predominantly form axospinous synapses in the lateral septal area, they terminate mainly on the dendritic shafts and soma of the vasopressin-receptive neurons. Of 720 vasopressin-immunoreactive terminals in the mediolateral part of the lateral septal area, 80% form synaptic contacts with dendritic shafts: 50% on small (distal) dendritic profiles and 30% on large (proximal) dendrites. Synaptic contacts between vasopressin-immunoreactive terminals and dendritic spines were not observed. The remaining 20% of immunoreactive boutons formed axosomatic synaptic contacts with a total of 58 neurons; 31% of these neurons exhibited somatic spines in the plane of the section analysed. Previous studies have demonstrated that in the lateral septal area vasopressin modulates the action of the excitatory amino acid-containing hippocamposeptal fibers, and also plays a role in the maintenance of long term potentiation evoked by fimbria-fornix stimulation. The convergent vasopressinergic and hippocampal input onto the same somatospiny neurons of the lateral septal area suggests that these neurons are targets of these physiological actions.

The roles that glutamate N-methyl- d -aspartate (NMDA) and dopamine D1-like and D2-like receptors play in the cortical neurotoxicity occurring in rats exposed to multiple doses of amphetamine (AMPH) for 2 days was evaluated. Neurodegeneration in rats that did not become hyperthermic during AMPH exposure was quantified by counting isolectin B4-labeled phagocytic microglia and Fluoro-Jade (F-J)-labeled neurons in the somatosensory parietal cortex, piriform cortex and posterolateral cortical amygdaloid nucleus (PLCo). The NMDA receptor antagonist, dizocilpine (0.63 mg/kg day) blocked AMPH-induced neurodegeneration in the somatosensory cortex. However, it did not affect degeneration in the piriform cortex and PLCo indicating that limbic degeneration was not NMDA-mediated. The dopamine antagonists, eticlopride (D2/3, 0.25 mg/kg day) and SCH-23390 (D1, 0.25 mg/kg day), blocked the stereotypic behavior and neurodegeneration in the somatosensory cortex. However, eticlopride had a lesser protective effect in the limbic regions. As well, the dopamine D2/D3 agonist quinpirole (1.5 mg/kg day) protected against cortical neurodegeneration when it was given during AMPH exposure and continued until sacrifice. The dopamine D1 agonist (SKF-38393, 12.5 mg/kg day) had no significant effect on neurodegeneration. These data indicate that there are significant differences in NMDA and dopamine D2 modulation of AMPH-induced neurodegeneration in the somatosensory cortex compared to the limbic cortices, and limbic cortical degeneration is not necessarily dependent on excessive stimulation of NMDA receptors as it is in the somatosensory cortex. Although excessive dopamine receptor stimulation during amphetamine exposure may trigger the neurodegenerative processes, continued D2 stimulation after AMPH exposure is neuroprotective in the cortex.

It was the objective of this retrospective study to analyse the causes of injury, surgical approaches, outcome, and complications in patients with vascular trauma and to report our experience with vein homografts for arterial reconstruction in the upper and lower limbs and cervicothoracic region in patients operated on over a period between 1981-2001.In 128 patients with peripheral arterial injuries the mechanism was direct penetration in 90 cases and blunt injury in 20 cases. In 4 cases a chronic damage (false aneurysm, AV fistula) was observed. Isolated vascular trauma was present in 97 patients (75.8%), 31 cases (24.2%) were aggravated by concomitant bone fractures, and nerve or soft tissue damage. Most frequently injured vessels were the superficial femoral (22.6%), crural (22.6%), and ulnar and radial (13.2%) arteries. 16 patients with penetrating cervicothoracic arterial injuries were registered during this period. 8 patients underwent emergency exploration and 8 patients angiography prior urgent exploration.Saphenous vein interposition grafting was applied with good results in 34 patients, polytetrafluoroethylene and Dacron grafts were used in eight cases, end to end anastomosis in 12 cases, venous bypasses in 5 cases, venous patches in 7 cases. 17 patients underwent arterial repair and 9 venous repair. Vein homografts as an arterial substitute were implanted in 8 patients. Five secondary amputations were performed and five patients died. The limb salvage rate was 95% and the primary patency rate of vein homografts was 75% (excluded primary amputations). The penetrating cervicothoracic injuries were all repaired with improvement in level of consciousness and neurological deficit when present.Most vascular injuries of the extremities can be managed successfully unless associated with severe concomitant damage of the bones, nerves and soft tissues. In the absence of suitable autologous vein grafts, homografts appear to be an interesting alternative for arterial repair. In penetrating cervicothoracic vascular injuries immediate operative repair offers the best chance of recovery.

Gene expression profiles in several brain regions of adult male rats were evaluated following a d -amphetamine (AMPH) exposure paradigm previously established to produce AMPH neurotoxicity. Escalating doses of AMPH (5–30 mg/kg) were given over the course of 16 h per day in an 18 °C environment for 2 days. This paradigm produces neurotoxicity but eliminates or minimizes the hyperthermia and seizure activity that might influence gene expression in a manner unrelated to the neurotoxic effects of AMPH. The expression of 1185 genes was monitored in the striatum, parietal cortex, piriform cortex and posteriolateral cortical amygdaloid nucleus (PLCo) using cDNA array technology, and potentially significant changes were verified by RT–PCR. Gene expression was determined at time points after AMPH when neurodegeneration was beginning to appear (16 h) or maximal (64 h). Expression was also determined 14 days after AMPH to find long-term changes in gene expression that might be biomarkers of a neurotoxic event. In the parietal cortex there was a two-fold increase in neuropeptide Y precursor protein mRNA whereas nerve growth factor-induced receptor protein I-A and I-B mRNA decreased 50% at 16 h after the end of AMPH exposure. Although these changes in expression were not observed in the PLCo, insulin-like growth factor binding protein 1 mRNA was increased two-fold in the PLCo at 16 and 64 h after AMPH. Changes in gene expression in the cortical regions were all between 1.2- and 1.5-fold 14 days after AMPH but some of these changes, such as annexin V increases, may be relevant to neurotoxicity. Gene expression was not affected by more than 1.5-fold at the time points in the striatum, although 65% dopamine depletions occurred, but the plasma membrane-associated dopamine transporter and dopamine D2 receptor were decreased about 40% in the substantia nigra at 64 h and 14 days post-AMPH. Thus, the 2-day AMPH treatment produced a few changes in gene expression in the two-fold range at time points 16 h or more after exposure but the majority of expression changes were less than 1.5-fold of control. Nonetheless, some of these lesser fold-changes appeared to be relevant to the neurotoxic process.

The dopamine-releasing and depleting substance amphetamine (AMPH) can make cortical neurons susceptible to damage, and the prevention of hyperthermia, seizures and stroke is thought to block these effects. Here we report a 2-day AMPH treatment paradigm which affected only interneurons in three cortical regions with average or below-average dopamine input. AMPH (six escalating doses/day ranging from 5 to 30 mg/kg for 2 days) was given at 17-18 degrees C ambient temperature (T) to adult male rats. During the 2-day AMPH treatment, peak body T stayed below 38.9 degrees C in 40% of the AMPH treated rats. In 60% of the rats, deliberate cooling suppressed (

Serum concentration of antibodies to C1q (C1qAb) has been reported to be elevated in a high percentage of patients with systemic lupus erythematosus (SLE). The associations of high C1qAb levels with different clinical manifestations and the activity of the disease, however, are not definitely understood.We measured the levels of IgG type C1qAb in the sera of 137 patients with SLE using an ELISA method.Serum concentrations of C1qAb were found to be higher (p0.0001) in SLE patients than in healthy controls. High titer (66 AU/ml) C1qAb was found in 40/137 (29.2%) SLE patients, and 4/192 (2.1%) healthy controls (p0.0001). A strong negative correlation (R = -0.4, p0.0001) between the age of the patients and the C1qAb titers could be detected. C1qAb levels in clinically active SLE patients significantly (p0.0001) exceeded those measured in the sera of patients in the inactive stage of the disease. A significant positive correlation was detected between C1qAb levels and the laboratory activity markers (anti-DNA, low C3 level) of the disease. We found a significant negative correlation between levels of C1qAb and a negative acute phase protein, alpha2-HS-glycoprotein. Renal involvement was present in 11/40 (27.5%) and 11/97 (11%) of the patients with high and low titers of C1qAb, respectively (p = 0.038). The prevalence of other organ manifestations was, however, the same in the patients with or without high titer C1qAb.These findings indicate that C1qAb measurement is a useful method for detecting the activity of SLE and predicting renal manifestations, but not other organ involvement in the disease.

The cellular and subcellular localization of muscarinic receptor proteins m1 and m2 was examined in the neostriatum of macaque monkeys by using light and electron microscopic immunocytochemical techniques. Double-labeling immunocytochemistry revealed m1 receptors in calbindin-D28k–positive medium spiny projection neurons. Muscarinic m1 labeling was dramatically more intense in the striatal matrix compartment in juvenile monkeys but more intense in striosomes in the adult caudate, suggesting that m1 expression undergoes a developmental age-dependent change. Ultrastructurally, m1 receptors were predominantly localized in asymmetric synapse-forming spines, indicating that these spines receive extrastriatal excitatory afferents. The association of m1-positive spines with lesion-induced degenerating prefronto-striatal axon terminals demonstrated that these afferents originate in part from the prefrontal cortex. The synaptic localization of m1 in these spines indicates a role of m1 in the modulation of excitatory neurotransmission. To a lesser extent, m1 was present in symmetric synapses, where it may also modulate inhibitory neurotransmission originating from local striatal neurons or the substantia nigra. Conversely, m2/choline acetyltransferase (ChAT) double labeling revealed that m2-positive neurons corresponded to large aspiny cholinergic interneurons and ultrastructurally, that the majority of m2 labeled axons formed symmetric synapses. The remarkable segregation of the m1 and m2 receptor proteins to projection and local circuit neurons suggests a functional segregation of m1 and m2 mediated cholinergic actions in the striatum: m1 receptors modulate extrinsic glutamatergic and monoaminergic afferents and intrinsic GABAergic afferents onto projection neurons, whereas m2 receptors regulate acetylcholine release from axons of cholinergic interneurons. J. Comp. Neurol. 434:445–460, 2001. © 2001 Wiley-Liss, Inc.

Estrogen receptors (ER) play a significant role in the development of some regions of the mammalian brain. Recently, ER-beta (ERbeta) mRNA and protein were shown to be expressed in the rat cerebellum. In the present study, the ontogeny of ERbeta protein expression was examined in the rat cerebellum during postnatal development. Western blot analysis indicated that a single ERbeta-like immunoreactive species of approximately 55 kDa was present in protein lysates prepared from the cerebella of female and male Sprague-Dawley rat pups. Immunocytochemical analysis of cerebellar sections from the midline vermis revealed that during development, the expression of ERbeta varied with age and cell-type, but not sex. In the developing cerebellum, highest levels of ERbeta-immunoreactivity (IR) were detected in neurons during neurite growth, and in some glia during migration. Throughout the first postnatal week, ERbeta-IR was localized to differentiating granule cells in the external germinal layer and to migrating glia. Differentiating granule cells expressed detectable levels of ERbeta throughout development. In Purkinje cells, ERbeta-IR was first detected on postnatal day 6 (P6), with peak intensities of immunostaining coinciding with the initiation of axonal and dendritic growth that occurs between P7 and P8. Expression of ERbeta-IR remained high during maturation of Purkinje cell dendrites, and then decreased to a lower level maintained in the adult. From the third postnatal week, ERbeta-IR was also detected in the later developing Golgi, stellate, and basket neurons. These results suggest that ERbeta may play a role in growth-related mechanisms during differentiation of cerebellar neurons and glia.

Soluble L-selectin (sL-selectin) concentrations were measured in paired samples of serum and cerebrospinal fluid by an ELISA method. Patients with several forms of multiple sclerosis (MS) and systemic lupus erythematosus with central nervous system involvement (SLE-CNS) were investigated. Elevated CSF sL-selectin concentrations were found in patients with SLE-CNS (7.62 +/- 3.31 ng/ml) and with relapsing-remitting form of MS (6.99 +/- 4.72 ng/ml) compared to the control group (4.00 +/- 0.95 ng/ml). The data presented suggest some similarities between inflammatory/immunological events in the central nervous system in patients with SLE-CNS and relapsing-remitting form of MS. Immunological heterogeneity in MS is suspected.

Interleukin (IL) 6 has an important role in the regulation of acute-phase proteins (APPs) during an acute-phase response. We studied IL-6 and other cytokines to determine if they regulate serum APP levels in the same way under the condition of the aberrant, long-lasting 'acute-phase response' that occurs in patients with chronic inflammation and cancer.Serum levels of nine positive APPs [CRP, SAA, C1-INH, Bf, C5, C8, C9, alpha 1-acidic glycoprotein (AGP) and haptoglobin] and two negative APPs [transferrin and alpha 2-HS glycoprotein (AHSG)] were measured using immunochemical methods in 59 multiple myeloma patients and in 72 healthy control subjects. Serum IL-6 and tumour necrosis factor (TNF) alpha levels were determined by bioassays.IL-6 was negatively correlated with five out of nine (C1-INH, C8, C9, AGP and haptoglobin) positive APPs but positively correlated with C-reactive protein (CRP). When patients with high and low IL-6 serum concentration were compared, CRP levels were higher, AGP and haptoglobin levels were lower in the high- than in the low-L-6 group, whereas no significant difference between the two groups was found in levels of the other positive and negative APPs. TNF-alpha levels were negatively correlated with transferrin and AHSG levels. No difference in the levels of positive APPs was observed between patients with low and high TNF-alpha serum concentration. By contrast, levels of both transferrin and AHSG were significantly lower in the high- than in the low-TNF-alpha group.These findings indicate that, except for regulation of the negative APPs by TNF-alpha, the mechanism of APP regulation is different under the conditions of the short-term and the chronic, long-lasting 'acute-phase reaction'.

Authors report a case in which relapsing polychondritis had been diagnosed two years before myelodysplastic syndrome developed and terminated in eosinophilic leukemia. The observation that relapsing polychondritis may precede myelodysplasia is not in concordance with some of the previous reports regarding relapsing polychondritis as a paraneoplastic phenomenon of myelodysplastic syndrome. The terminally developed eosinophilic leukemia is not supposed to be a blastic phase of the underlying myelodysplasia, much rather a second malignant process. This opinion may be confirmed by the early presence of blast cells in the myelodysplastic process without eosinophilia. It seems interesting to note that both our patient and his daughter suffered from diseases of autoimmune origin: acquired vitiligo and subacute cutan lupus erythematodes, respectively.

It has been shown that sexual dimorphic morphology of certain hypothalamic and limbic areas underlie gender-specific sexual behavior and neuroendocrine mechanisms. The key role played by locally formed estrogen in these developmental events has been revealed during a critical perinatal period. In this study, we aimed to document the presence of estrogen-synthetase (aromatase)-immunoreactive elements in the involved limbic system and hypothalamus of the developing rat brain. On postnatal day 5, animals of both sexes were perfusion-fixed, and sections from the forebrain and hypothalamus were immunolabelled for aromatase using an antiserum that was generated against a 20 amino acid sequence of placental aromatase. Aromatase-immunoreactivity was present in neuronal perikarya and axonal processes in the following limbic structures: the central and medial nuclei of the amygdala, stria terminalis, bed nucleus of the stria terminalis (BNST), lateral septum, medial septum, diagonal band of Broca, lateral habenula and all areas of the limbic (cingulate) cortex. In the hypothalamus, the most robust labelling was observed in the medial preoptic area, periventricular regions, ventromedial and arcuate nuclei. The most striking feature of the immunostaining with this antiserum was its intracellular distribution. In contrast to the heavy perikaryal labelling that can be observed with most of the currently available aromatase antisera, in the present experiments, immunoperoxidase was predominantly localized to axons and axon terminals. All the regions with fiber staining corresponded to the projection fields of neuron populations that have previously been found to express perikaryal aromatase. Our results confirm the presence of aromatase-immunoreactivity in developing limbic and hypothalamic areas. The massive expression of aromatase in axonal processes raises the possibility that estrogen formed locally by aromatase may not only regulate the growth, pathfinding and target recognition of its host neuronal processes, but may also exert paracrine actions on structures in close proximity, including the target cells.

Intraneuronal production of estradiol from testosterone has been shown to play a pivotal role in gender-specific brain development of most vertebrates, and to participate in numerous functions of the adult central nervous system. Previous biochemical and morphological approaches demonstrated that estrogen synthetase (aromatase) is present in specific limbic and hypothalamic structures. On the other hand, less attention has been paid to revealing its subcellular distribution. The possibility of aromatase presence in axonal processes has been indicated by recent biochemical and morphological observations suggesting new insights for the role of aromatase in neural functions. The objective of the present study was to provide morphological evidence for the subcellular location of aromatase in neurons of different vertebrate species including Japanese quail, rat, monkey, and human. Immunocytochemistry using a purified polyclonal antiserum against human placental aromatase localized immunoreactivity to hypothalamic and limbic cell groups in all of these species. Light and electron microscopic examination of vibratome sections revealed the presence of aromatase immunoreactivity throughout the neuronal perikarya, including dendrites and axonal processes. In each species there were numerous boutons which contained labeled small clear synaptic vesicles. Many of these axon terminals formed synapses with immuno-negative and immuno-positive dendrites and perikarya. This study furnishes the first immunolocalization of aromatase in the brains of two primate species, humans and monkeys. The provision of further evidence for estrogen synthesis in axons and axon terminals may help resolve apparent differences between the measurement of aromatase activity and the lack of aromatase-immunopositive cell bodies in previous studies. The present findings may be coupled with recent evidence regarding the molecular biology and the diversity of functional properties of P450 aromatase to indicate previously unexpected effects of brain aromatase at the synaptic level.

The electrophysiological observations that substance P administration to the lateral septal area elicits both excitatory and inhibitory responses, together with earlier reports on the multiple sources of substance P innervation of the septum, implies that these axons with distinct origins have different functions. This prompted us to examine the origin and neurochemical character of substance P afferents to the lateral septal area. Chronic surgical isolation of the septum from its ventral afferents and retrograde tracer experiments using wheat germ agglutinin-conjugated horseradish peroxidase, both followed by an immunostaining for substance P, were employed to elucidate the origin of these axon terminals. In order to assess the possible co-existence of substance P with other neurotransmitter substances in the parent cells of the septopetal projections, co-localization studies for substance P and choline acetyltransferase, as well as substance P and GABA, were performed. The comparative distribution of substance P fibers and septal calbindin-containing neurons was also investigated using correlated light and electron microscopic double immunostaining. The results are summarized as follows: (i) the substance P innervation of the lateral septal area derives from several hypothalamic nuclei (including the lateral and lateroanterior hypothalamic area, tuber cinereum and ventromedial hypothalamic nucleus) and tegmental nuclei (the majority of fibers from the laterodorsal and a few from the pedunculopontine tegmental nucleus), as well as intrinsic septal cells; (ii) the septopetal substance P fibers of tegmental origin are cholinergic; intraseptal substance P neurons located in the dorsolateral part of the lateral septum also contain GABA, while substance P neurons seen on the border between the medial and lateral septal area and septopetal hypothalamic substance P cells do not contain GABA or acetylcholine; (iii) substance P fibers from pericellular baskets around calbindin-containing lateral septal neurons with a high degree of selectivity; (iv) approximately 90% of the entire calbindin cell population are postsynaptic targets of substance P axons; (v) their terminals contact the soma and the dendrites of these cells, among them the somatospiny neurons; and (vi) the extrinsic substance P boutons establish asymmetric, while the intrinsic substance P axon terminals form symmetric membrane specializations. Because neurons in the lateral septal area receive hippocampal input and project massively to hypothalamic areas, the different types of substance P input on these neurons can modify the information flow arriving from the hippocampus to diencephalic brain structures at the level of the lateral septal area.

The author summarizes the newest results and knowledges which are related to the pathogenesis of polyarthritis chronica progressiva. Pursuing the composed target the feasible roles of gamma delta TcR T lymphocytes and stress proteins as well as that of CD5 positive B lymphocytes and IgG-glycosylation status are discussed. Reaching the end of the paper it is established that the solution of etiology and pathogenesis of polyarthritis chronica progressiva, in spite of the increasing theoretical knowledge, awaits for further extensive research work.

In the lateral septal area (LSA), both inhibitory and excitatory dopamine (DA) actions, as well as hypothalamic and midbrain DA efferents, have been described. Some neurons of the hypothalamic and midbrain DA systems contain somatostatin (SOM) or neurotensin (NT), and, in the LSA, the distribution of fibers containing these peptides overlaps with DA fibers. These data prompted us to test for the presence of SOM and NT in LSA dopaminergic axon terminals of hypothalamic and midbrain origins. To verify the origins of SOM and NT innervation of the LSA, the retrograde tracer horseradish peroxidase conjugated with wheat germ agglutinin (HRP-WGA) was injected into the LSA, and alternate brain sections were immunostained for SOM, NT, or tyrosine hydroxylase (TH) in group 1 rats. Numerous retrogradely labeled neurons were found immunopositive for SOM in the periventricular and basolateral hypothalamus, many HRP-WGA labeled cells contained NT immunoreactivity in the ventral tegmental area, and TH-immunoreactive retrogradely labeled neurons were observed in both brain areas. In a new approach, the presence of these peptides in dopaminergic boutons was assessed by combining peptide immunocytochemistry with acute 6-hydroxydopamine (6-OHDA) induced lesioning of DA cell groups. These groups of rats were treated with desipramine to protect the noradrenergic fibers, and 45 min later 1 microgram 6-OHDA (in 0.5 microliter saline) was unilaterally injected into the periventricular hypothalamus (group 2) or the ventral tegmental area (group 3). After 48 h the rats were killed and alternate septal sections of both groups were immunostained for TH, SOM, or NT. On the operated side of the LSA in both groups, electron microscopy revealed numerous axon terminals that were immunopositive for TH and contained autophagous cytolysosomes, an early sign of catecholamine fiber degeneration induced by 6-OHDA. In group 2, phagosome-containing boutons were found immunopositive for SOM, but not for NT; vice versa, in group 3, only NT-positive degenerated boutons were detected. SOM- and NT-positive degenerated axon terminals in both groups formed synaptic contacts with LSA neurons, in particular with somatospiny cells. On the contralateral side of the LSA, all of the axon terminals were intact. It has been shown that SOM exerts an inhibitory action, whereas NT has an excitatory effect on limbic area neurons. Thus, the results implicate that the differential peptide content of dopamine fibers marks their functional differences. It appears that LSA neurons receive double innervation from an inhibitory "somatostatinergic" DA system of the hypothalamus, and from an excitatory "neurotensinergic" DA system of the midbrain.

This study deals with the synaptology, morphologically identified postsynaptic targets, and origin of somatostatin (SOM) fibers in the rat lateral septal area (LSA) with special reference to those forming pericellular baskets. Septal vibratome sections were immunostained for SOM-14 in 3 experimental groups: control animals, rats subjected to a chronic transection of the ascending afferents to the septum, and animals with acute fimbria-fornix lesion. Light microscopy revealed that the SOM-immunoreactive fibers form pericellular baskets predominantly in the intermediate and ventral parts of the caudal half of the LSA. Electron microscopic analysis showed that the somatospiny neurons are postsynaptic targets of these pericellular baskets. Eight days after a unilateral cut placed at the ventral border of the septum, virtually all SOM-immunoreactive axon terminals disappeared from the ipsilateral intermediate and ventral LSA, and they were substantially reduced in the dorsal LSA. However, in these rats SOM-positive neurons could be observed in the LSA on the lesioned, but not on the contralateral side. Furthermore, on the lesion side of the anterior periventricular hypothalamus an increase was detected both in the number and the intensity of immunostaining of SOM-positive neurons. Thirty-six h following a unilateral transection of the fimbria-fornix, the SOM-immunoreactive axon terminals in the LSA remained intact; only immunonegative degenerated hippocamposeptal boutons were detected forming synaptic contacts with somatospiny neurons. Axosomatic synapses of SOM-positive boutons regularly appeared at the neck of somatic spines which were postsynaptic to degenerated hippocamposeptal fibers. The results indicate that the septal SOM fibers are of multiple origin. Those forming pericellular baskets in the LSA originate in ventral extraseptal, probably periventricular hypothalamic areas. SOM fibers scattered in the dorsal LSA are most likely processes of local SOM neurons. The accumulation of immunoreactive SOM in some cells of the undercut septum is a sign of axonal lesion, indicating that these neurons project outside the septum. The SOM innervation of somatospiny neurons which also receive hippocampal input and have been reported to contain γ-aminobutyric acid (GABA) may be a morphological substrate of the SOM-related disinhibition in the LSA.

Authors investigated the in vitro effect of alpha 2-HS-glycoprotein on the lymphoblastic transformation and interleukin-2 production. The observations were made on the peripheral blood lymphocytes of healthy persons, on those of women taking oral contraceptives and on those of patients with systemic lupus erythematosus. The alpha 2-HS-glycoprotein inhibited the mitogen responses both in physiologic and in the halved concentrations. The inhibition of the interleukin-2 production seemed less remarkable.Whether blast transformation of human lymphocytes from healthy volunteers, women taking oral contraceptives, and women with systemic lupus erythematosus (SLE) would be affected by alpha2-HS-glycoprotein (A2HSGP) was explored by incubating lymphocytes stimulated by phytohemagglutinin or pokeweed mitogen with or without the glycoprotein. A2HSGP did indeed inhibit blast transformation in these cell populations. Release of the peptide interleukin-2 into the culture medium was also inhibited to a lesser extent, slightly more so in cells from healthy persons, and slightly less so in women with SLE. The mechanism and biological significance of this observation is unexplained.

Serum alpha 2HS-glycoprotein (A2HSG) concentrations of 63 patients with systemic lupus erythematosus (SLE) were determined, and found to be significantly low compared to those of 59 healthy blood donors. The diminution of serum A2HSG concentration was proportional to the degree of activity of SLE, and was not influenced by secondary infections. There was a statistically significant positive correlation between serum A2HSG and the C3 complement component levels. A negative correlation between serum A2HSG and IgG, IgA concentration and anti-DNA activity was observed. Serum A2HSG was significantly low in cases of positive for the following laboratory parameters: anti-nuclear antibodies, circulating immune complexes and LE cell phenomenon. We found no correlation between serum IgM concentration, cryoglobulins, latex agglutination and serum A2HSG levels. The unusually good negative correlation between A2HSG pathogenetical role of this glycoprotein in SLE. The determination of A2HSG concentration may be of clinical importance in SLE.

As a result of a single intraperitoneal treatment with 1 mg/ml of Mannozym (M) the plasma fibronectin (FN) level was significantly increased both in germfree (Gf) and conventional (Cv) mice, and though it started from a lower value in Gf mice, it rose to a similar level to that of Cv animals. The maximum of FN level was observed on the first day after treatment both in Gf and Cv mice. It had returned to normal by the 7th day in Cv mice, but it was higher in Gf mice as compared to untreated controls even on the 14th day. Thus the increase of FN level was of higher degree and longer duration following M treatment in Gf mice than in Cv animals. In treated Gf mice the plasma FN concentration was in the same range as in untreated Cv mice even at the termination of experiment. Both in Gf and Cv mice, there was a relative spleen weight increase, the degree of which was similar, but the duration was longer in Gf mice than in Cv ones.

The feed of weaned piglets of Hungarian Large White X Duroc and Dutch Landrace X Duroc genotype was supplemented with 0.5 mg selenium, 50, 100 or 150 mg vitamin E, and 2.5 or 5 mg riboflavin per kg. Feed supplementation enhanced the cytotoxic reaction and elevated the antibody titres produced against purified horse gamma globulin antigen. However, as compared to the control the differences were not significant. Feed supplementation exerted a beneficial, though varying, influence on the indices of cell-mediated immunity. The proportion of rosette-forming cells and blastogenic transformation induced by specific (horse globulin) and nonspecific (phytohaemagglutinin, PHA) mitogens underwent the most expressed and most significant increase in pigs fed 5 mg selenium, 100 mg vitamin E and 5 mg riboflavin per kg of feed. On the other hand, feed supplementation failed to enhance the responsiveness to intradermal PHA (type IV allergic reaction).

Plasma fibronectin concentration was determined by electroimmunodiffusion and laser nephelometry in 40 healthy persons and 321 patients with myelo- and lymphoproliferative diseases and other malignancies. Decreased fibronectin concentrations were found in patients with leukemia, Hodgkin's and non-Hodgkin's lymphomas, myelofibrosis, polycythaemia rubra vera and angioimmunoblastic lymphadenopathy. Elevated fibronectin level was detected in patients with multiple myeloma. In patients with cancer of the lung, stomach and colon, fibronectin level was found in the normal range. Decreased fibronectin concentration was observed in patients with cancer of the breast and prostate. Lower plasma fibronectin concentrations were detected in all groups of patients with infectious septical complications as compared to the patients without infections.

Changes of T and B lymphocyte populations have been studied in chronic active hepatitis and liver cirrhosis. It was found that E rosette-forming T lymphocyte counts were significantly lower in this group of diseases than in the controls. The B cell count did no change significantly as compared to the controls. Incubation with levamisole was followed by a significant increase in active E rosette-forming and total T-cell counts. The authors have appraised the experimental results by comparison to published findings.

The effect of N-acetyl-4-6-(p-oxy-benzylidene)-D-glycosamine on Na235SO4 incorporation has been studied in vitro. The 0.25-2.0 mmoles of the compound significantly inhibited the Na235SO4 incorporation of granulomatous tissue. The inhibitory effect proved to be dose dependent.

The number of circulating total lymphocytes and the number of B and T lymphocytes from the blood were estimated in 22 patients with biopsy-verified chronic active liver disease. The total number of lymphocytes and the number of rosette-forming cells (T lymphocytes) were decreased in patients with liver disease. The number of IgG,- IgA- and IgM-bearing lymhocytes (B cells) did not change significantly while the number of null cells increased in patients vs. a control group.

Chronic hepatitis was diagnosed on the basis of biochemical, immunological and morphological criteria in 153 cases. On the evidence of observations for a mean period of four years the prognosis of chronic persistent hepatitis is regarded as favourable, no progression to chronic aggressive hepatitis or to cirrhosis having been observed in any of the cases. On the other hand, chronic aggressive hepatitis was found to progress to cirrhosis in 12 out of 65 cases. Cirrhotic transformation was more frequent in hyperactive processes (8 out of 25 cases). The sera of patients with primary hepatocellular carcinoma showed low immunoglobulin concentrations, with increased coeruloplasmin and reduced transferrin levels.

The effect of alcohol ingestion on the prognosis of chronic liver diseases was studied'n. Among 880 patients with chronic liver diseases the percentage of alcoholics was high, 33%, compared to 3.2% in the control group. The period of survival time is decreased by one half in patients with regular alcohol ingestion. According to 'in vitro' investigations they report that in chronic active hepatitis the low Et-rosette forming cell number is further decreased after incubation with alcohol, and ethanol inhibits the immune-stimulating effects of levamisole on Ett-rosette forming cells.

Fibronectin was detected by immunofluorescence on the surface of one fraction of separated normal peripheral blood lymphocytes using FITC-conjugated anti-human fibronectin antibodies. Approximately one fifth of isolated B cells and 7% of O cells contained surface bound fibronectin but T cells failed to stain. There were no detectable free receptors for fibronectin on the surfaces of the lymphocytes in the different subsets isolated from healthy controls as studied using FITC-labelled purified fibronectin. The per cent of B and O cells bearing surface bound fibronectin was markedly decreased in patients with acute and chronic lymphocytic leukemias and non Hodgkin's lymphoma, only 1-4% of B and 1-2% of O cells stained with FITC-labelled antifibronectin immunoglobulins. FITC-conjugated fibronectin was not bound to the different lymphoblasts isolated from patients with leukemia and lymphoma. Treatment of cells with trypsin and EDTA removed fibronectin bound to the cell surfaces. Fibronectin attached to the surfaces of lymphocytes may have an immunoregulatory function.

The concentration of IgA, IgG, IgM, coeruloplasmin, alpha-2-macroglobulin, beta-1-C-globulin, transferrin as well as of alpha-2-haptoglobin, orosomucoid and haemopexin has been studied in the serum of patients with disseminated lupus erythematosus, chronic progressive polyarthritis and immunopathological syndromes. The concentration of the carbohydrate components bound to serum proteins was also determined. In disseminated lupus erythematosus, the concentration of IgA, IgG, IgM, coeruloplasmin, and hexose, hexosamine and sialic acid bound to proteins, as well as of seromucoid was increased. In chronic progressive polyarthritis the IgA, IgG, IgM, coeruloplasmin, alpha-2-macroglobulin and beta-1-C-globulin concentration and the carbohydrate components bound to serum proteins rose. In immunopathological syndromes, too, the serum IgA, IgG, IgM, coeruloplasmin, and protein-bound hexosamine and sialic acid levels were increased. Under the effect of treatment, the elevated glycoprotein concentrations usually decrease, but the level of carbohydrate components bound to serum proteins hardly changes. The pathogenic and diagnostic significance of serum glycoproteins is discussed.

Serum cholesterol, triglyceride, total lipids and the lipoprotein pattern were studied in 169 cases chronic liver disease confirmed by biopsy. On the ground of the immunological and morphological results the patients were classified into five groups. In chronic persistent hepatitis no significant abnormality was found. In chronic aggressive hepatitis and in cirrhosis of the liver the serum cholesterol level was significantly reduced. In fatty infiltration of the liver the serum cholesterol, triglyceride and total lipid concentrations were significantly increased, as compared with the normal values and with the figures obtained in the cases of chronic inflammatory liver disease. In the cases of cirrhosis with additional diabetes the lipid values were likewise increased. In chronic aggressive hepatitis and in cirrhosis of the liver the levels of pre-beta and alpha lipoprotein were decreased, in fatty infiltration of the liver those of beta and pre-beta lipoprotein were increased.

Lower plasma fibronectin (FN) levels were detected in C3H germfree (Gf) mice as compared to conventional (Cv) ones. FN levels increased with aging both in Gf and Cv mice. Stimulation of the immune system with live Newcastle disease virus (NDV) vaccine increased the FN levels in young germfree mice, but had no effect in conventionals.

The sera of patients with IgG-myeloma, IgM-macroglobulinaemia and with processes accompanied by high polyclonal Ig-levels were measured by radial immunodiffusion for immunoglobulins, coeruloplasmin, alpha-2-macroglobulin, C3-component, transferrin, alpha-1-antitrypsin, alpha-2-haptoglobin and haemopexin. In the presence of typical changes in the immunoglobulin concentrations, elevation of alpha-2-macroglobulin was found only if polyclonal Ig was increased. The concentration of the C3 component was reduced only in IgM macroglobulinaemia, that of transferrin only in IgG-myeloma, that of alpha-2-haptoglobin only in IgM-macroglobulinaemia and in case of increased polyclonal Ig.

In liver biopsy specimens of 45 patients with chronic persistent hepatitis, chronic aggressive hepatitis and liver cirrhosis the number of lymphoid cells and fibroblasts as well as in the sera of the same patients the concentration of IgG, IgA, IgM, alpha-2-macroglobulin and coeruloplasmin have been studied. The number of lymphoid cells and fibroblasts, was significantly elevated in chronic aggressive hepatitis and liver cirrhosis; a close correlation could be demonstrated between the number of the lymphoid cells and the IgG concentration; the serum alpha-2-macroglobulin level changed parallel to the number of liver fibroblasts in chronic aggressive hepatitis and liver cirrhosis.

Peripheral blood lymphocytes and the "in vitro' changes of the T-cell count after levamisole treatment were studied in twenty-two patients with systemic lupus erythematosus and in eighteen normal controls using E-rosette assay to quantify T-cells and EAC-assay to quantify B-lymphocytes. T-cell count was significantly lower in the patient group compared with the control group. B-cell numbers did not change significantly. The decrease of the T-cell count was accompanied by an increase of the null-cell number in patient groups. The levamisole treatment "in vitro' produced an increase in the active-T and total T-lymphocyte counts and led to the decrease of null-cell numbers. These data confirm the available evidence that levamisole "in vitro' may induce the maturation of immature T-lymphocytes in SLE.

In the serum of 27 patients with malignant lymphogranulomatosis the authors determined the serum level of glycoproteid-carbohydrate components (hexose, hexosamine, sialin acid, and seromucoid) and the concentration of 11 different glycoproteids. In the early stage of the disease the immunoglobulin level is moderately increased in the serum, whereas a diminution can be observed in stage IV. The concentrations of ceruloplasmin, alpha-2-macroglobulin and orosomucoid were already increased significantly in stage III. The increase did not continue in stage IV. In the final stage of the disease the concentrations of alpha-1-antitrypsin and haemopexin turned out to be increased considerably. A significant decrease in the transferrin level could be registered in stage III with this diminution also continuing in the further course. Changes of beta-C-globulin and haptoglobin concentrations could not be evaluated statistically. The content of carbohydrate components in the glycoproteids will already increase in the early stage of the disease with this increase continuing in the further course. Among histological types there was a more significant increase of immunoglobulins in those forms rich of lymphocytes.