Search

Mammalian choroid plexuses (CPs) are vascularized structures involved in numerous exchange processes that supply nutrients and hormones to the brain, and that remove deleterious compounds and metabolites from the brain. Studies in the adult Mediterranean buffalo have investigated the morphology of CPs using histochemical and immunohistochemical techniques. To date, however, there have been no studies conducted on ruminants regarding this removal process which serves to repair functional vascular damage in the CPs. Each of these vascular repair processes is a very complex and none of these has not yet been completely understood. Then, the aim of the present study is to investigate the morphological processes during angiogenesis in the CPs of healthy adult buffaloes, utilizing transmission electron microscopy (TEM), scanning electron microscopy (SEM), and immunogold-labeling SEM analysis (biomarkers: angiopoietin-2 [Ang-2], vascular endothelial growth factor receptor-3 [VEGFR-3], and CD133). At TEM, the inner surface of the blood capillaries sometimes showed pillar-like cells, which in contact with endothelial cells formed prominences, which in turn formed neo-blood capillaries. With immunogold-labeling SEM analysis, the CP blood capillaries showed Ang-2 and VEGF-3, respectively, in positive particles and spheroid formations. In addition, the external surface of the blood capillaries showed spheroid formations that originated from the neo-vascular capillaries whose terminals formed a capillary network, positive to CD133. On the basis of these results, the following hypothesis can be made, namely, that these CPs are vascular structures which play a fundamental role in maintaining brain homeostasis and self-repairing of functional vascular damage, independently of the presence of rete mirabile in this species. Microsc. Res. Tech. 75:1104–1112, 2012. © 2012 Wiley Periodicals, Inc.

1998).Since the lingual papillae of the domestic ruminant show an extreme variety of shape, webelieved it interesting to extend the study to innervation of the lingual papillae of the waterbuffalo, analyzing the morphological, structural, and immunohistochemical characteristicsof the gustatory papillae.MATERIALS AND METHODSThe study was performed on 25 water buffalo fetuses of both sexes, from 5 to 60cm inlength, and originating from a local slaughter-house. The fetus length, from the sinciput tothe tail root, was utilized as a development index. The fetuses were subdivided into fivegroups in relation to their length (5–7cm; 10–15cm; 20–25cm; 30–35cm; 40–60cm). Foreach group samples were prepared for scanning electron microscopy (SEM), transmissionelectron microscopy (TEM), light microscopy (LM), and the simple immunohistochemicalmethod.The TEM samples were treated by the usual techniques.

The buffalo is one of the few domestic animals that has a seasonal mating cycle, influenced by the photoperiod. It is known that the photoperiod regulates gonadal function probably via the pineal and/or hypothalamus-pituitary axis. Moreover, the hypothalamus (melatonin) and gonads influence the production of the signaling transmitter nitric oxide (NO), suggesting that the NO may have an important role in the regulation of gonadotropin-releasing hormone secretion. This further suggests the hypothesis that NO in the epididymis has an important role in the maturation of spermatozoa and their motility and posterior fertilization capacity. The aim of the present study is to investigate the seasonal variations in the morphology of the epididymis by means histochemical and immunocytochemical techniques. We used the NADPH-d, nitric oxide synthase (NOS) I and NOS III to clarify the relationship between epididymis function and NO signaling activity. The results of this work show that NO is present in the caput of epididymis during short photoperiods, i.e., periods of maximum gonadal activity (winter) and absent during long photoperiods, i.e., periods of gonadal regression according to the previously described role of NO in spermatozoa capacitation and motility in the caput epididymis. © 2012 Elsevier Inc. All rights reserved.

A group of 46 farm workers (32 men), affected by a recurrent “occupational disease of undetermined origin”, underwent an immunologic investigation with a Tetranychus urticae (TU) whole-body extract (TU-WBE) prepared in our laboratory. The patients suffered from seasonal attacks of rhinitis, during the summer and autumn periods, when working in open fields (30 subjects) or in greenhouse flower cultivation (16 subjects). In most patients, rhinitis was associated with bronchial asthma (16 subjects), urticaria (14 subjects), or both (three subjects). Allergic alveolitis or other common allergic diseases had been excluded, and a diagnosis of “occupational disease of undetermined origin” had been made before by other medical centers. Ten healthy farm workers and 10 atopic townsmen were chosen as control groups. An in vivo and in vitro diagnostic trial by skin prick testing (SPT) and serum specific IgE dosage with TU-WBE were done in all subjects. Thirty-six patients (78%) were found to be positive to both SPT and the IgE enzyme allergosorbent test (EAST), with a good correlation between IgE serum levels and cutaneous wheal size. Control groups did not show any reaction. The IgE-EAST homologous inhibition test was positive. The IgE-EAST cross-inhibition test excluded cross-reactivity between TU and Dermatophagoides pteronyssinus. The TU-exposure test was positive for the 36 patients with TU-WBE-specific IgE. Three patients who were negative for TU-WBE-specific IgE reacted to the TU-exposure test; in these patients, the scratch-chamber test (Finn chamber) with eggs and droppings from TU was positive. We suggest the possibility of a TU allergy also in those subjects in whom clinical symptoms can be attributed to other common allergens. Astarita C, Franzese A, Scala G, Sproviero S, Raucci G. Farm workers’occupational allergy to Tetranychus urticae: clinical and immunologic aspects.

The reticular groove mucosa of adult cattle, buffalo and sheep was investigated by histochemical and immunocytochemical techniques. Intense NADPH-d staining was observed in the folds of the epithelium mucosa and at the bottom of the reticular groove in all domestic ruminants studied. The NADPH-d staining showed that the innervations of the tunica muscularis of the reticular groove lip were composed of nerve corpuscles, nerve fibres and nerve cells of the mucosa epithelium. SEM analysis showed an intense nitric oxide synthase (NOS) I immunoreactivity in deep and medium cellular layers. It is interesting to note that the same morphologies were observed in samples of the mucosa epithelium, and of the tunica muscularis processed by NADPH-d and in those processed by immunogold techniques. This study has demonstrated that nitric oxide (NO) is involved in the rumination activity and that it plays a double role in this activity in the reticular groove of all domestic ruminants studied: (1) NO plays a role similar to the one it has in the mucosa epithelium of all the other compartments of the ruminant forestomach, (2) The lip sections of the reticular groove has shown abundant innervations that may indirectly coordinate and control the forestomach motility through the direct activation of the nitrergic (nitroxidergic) nerve cells and nerve fibres.

The choroid plexuses (CPs) in mammals produce the cerebrospinal fluid (CSF). In the literature, the morphology of CPs and the process that regulates the production of CSF are virtually nonexistent for domestic ruminants. Thus this study has two aims: 1. to investigate the morpho-structure of the buffalo CP microvasculature utilizing light microscopy (LM), scanning electron microscopy (SEM) and transmission electron microscopy (TEM) techniques, and 2. to investigate the relationship between the blood vessels and both the elongated cells and the cells with multiple protrusions located in the CPs. SEM and TEM analyses of the CPs from buffalo brain showed morphological and structural features similar those reported in other mammalian species. Moreover the blood microvasculature is the major component responsible for the formation of the CSF, secreted by the encephalic CPs. In addition the chemical composition of this fluid depends on several morpho-functional characteristics of the vascularization of the CPs. These characteristics are as follows: two shapes of the vascular organization: lamina-like and ovoid-like elongated cells of the CPs, which connect the ventricular cavities to the blood capillaries; and the CP capillaries have diverse forms. In the present study the employment of NADPHd and NOS I was taken as indirect evidence for the presence of NO for investigation their specific role in CPs. Then NOS I immunoreactivity is found in the walls of CP blood vessels demonstrating indirectly the presence of NO with a vaso-dilatatory and autoregulation function of vascular tone by cholinergic nerve stimulation of blood vessel smooth muscle. Microsc. Res. Tech. 74:67-75, 2011. © 2010 Wiley-Liss, Inc.

The forestomach plays an important role in the digestion physiology of ruminants. The aim of this study is to clarify the morpho-functional role of the mucosa in each of the three compartments of the forestomach in three domestic ruminants species, viz cattle, buffalo and sheep, by means of structural, histochemical and immunocytochemical methods, including transmission electron microscopy, light microscopy and scanning electron microscopy. These methods were chosen to demonstrate the indirect evidence for the presence of nitric oxide (NO) employing NADPHd and nitric oxide synthase I (NOS I). The various cell layers of the forestomach epithelium are described and illustrated in detail. An intense NADPHd staining was observed in the granulosa, spinosa and basal layers of the epithelium, in particular in the cytoplasm over the nucleus. NOS I immunoreactivity was found in all specimens of the forestomach mucosa. The results of this study might reflect a possible role of NO in delaying the onset of cellular apoptosis in the forestomach mucosa of the domestic ruminants, by playing a role in the production of cell energy.

The hypothalamic peptide orexin A, deriving from the proteolytic cleavage of the precursor molecule prepro-orexin, has a wide range of physiological effects including the regulation of feeding behaviour, neuroendocrine functions, sleep-wake cycle, and energy homeostasis. Lowered excretion of orexin A into the cerebrospinal fluid (CSF) plays a pathological role in animal and human narcolepsy. Altered levels of orexin A into the CSF have been also found in numerous disorders of the central nervous system, including Parkinson's and Huntington's disease, dementia, and depressive disorders. While the localization of orexin A and its receptor 1, OX(1), has been elicited in many regions of the mammalian brain and in peripheral organs, there are no information on the expression of the neuropeptide and its receptor 1 in the choroid plexuses (CPs) producing the CSF. In this study, we investigated the expression of orexin A and OX(1) in the CPs from the brain of an adult mammalian species, Bubalis bubalis, by immunogold-labelling in scanning electron microscopy. Both orexin A and OX(1) immuno-reactivity appeared to be widely distributed on the surface of choroid epithelium. Interestingly, a marked orexin A labelling was detected in the areas surrounding the CP blood capillaries. The expression of prepro-orexin and OX(1) mRNA transcripts of 200 and 300 bp, respectively, was assessed in the CPs by reverse-transcription polymerase chain reaction, while Western blotting analysis confirmed the presence of these two proteins in the tissue. Our findings provide the first evidence for orexin A and OX(1) expression in the CPs from mammalian brain, and suggest that the levels of orexin A into the CSF are probably regulated by CP activity. PMID: 19250669 [PubMed - indexed for MEDLINE]

Choroid plexuses (CPs) play pivotal roles in a wide range of processes that establish, survey, and maintain the biochemical and cellular status of the central nervous system. Mammalian CPs contain a very high density of serotonin receptors, and serotonin has been shown to affect CP functions. The serotonin transporter (SERT) regulates the entire serotonergic system, including serotonin receptors by means of modulation of serotonin concentration in the extracellular fluid. In this study, the expression of SERT in the CPs from the brain of a mammalian species, Bubalis bubalis, was established. By immunogold labeling in scanning electron microscopy, SERT immunoreactivity was found to be localized on the apical surface of the choroid epithelium. In particular, SERT positivity was detected on the apical portion of villi, and both on the membrane and in the cytoplasm of grouped cells on the surface of the choroid epithelium. Significantly, no SERT was detected in blood vessels irrigating the CPs. The expression of SERT mRNA transcripts of 440 bp in the CPs was detected by reverse-transcription polymerase chain reaction, and Western blotting analysis revealed the presence of three isoforms of the protein with molecular masses of approximately 70, 80, and 140 kDa, respectively, probably corresponding to differently glycosylated SERT. Our findings provide the first report of SERT detection in the CPs of buffalo brain and indicate that this protein is locally synthesized from the choroid epithelial cells. We suggest that SERT might have an important role in mammalian CPs, possibly regulating the serotonin flow between brain and rest of the body.

Choroid plexuses (CPs) play pivotal roles in many processes that establish, survey, and maintain the biochemical and cellular status of the central nervous system (CNS). Changes in the anatomy and physiology of CPs have been linked to several CNS diseases. However, CP structure and function are not definitely known. Here, we report structural and functional features of choroid epithelium from buffalo brain never described before. Mixed with common epithelial cells, two novel cell types were identified by scanning and transmission electron microscopies. The first peculiar cells showed a globular apical portion projecting into the ventricular cavities, and a basal peduncle in direct contact with blood capillaries underlying the epithelium. The second type of cells resulted to be formed by a globular body from which depart numerous processes; these cells, localized deeply in the choroid epithelium, strictly contact neighboring epithelial cells. No synaptic contacts were detected between these cell populations and common epithelial cells. To gain some insight into the functional properties of choroid cells, NADPH diaphorase (NADPHd) and neuronal nitric oxide synthase (nNOS) activities were evaluated. Of interest, whereas a strong NADPHd activity was detected in all cell types of choroid epithelium, nNOS was only detected in the first type of peculiar cells. The presence of nNOS in the CPs was confirmed by Western blotting. These results suggest that nitric oxide may serve as a signal for the regulation of CP multiple functions. Copyright 2007 Wiley-Liss, Inc.

The microcirculation of the foetal kidney was studied in the buffalo using light (LM) and scanning electron microscopy (SEM). The primordial glomerules originated from the peripheral zone of the metanephros at the stage of 8 cm CRT. The glomerular capillaries started to differentiate at the stage of 10-15 cm CRT. They were sparse and showed a few primordial pores. In addition, they began to make contacts with primordial podocytes. At the stage of 40-60 cm CRT, the renal microcirculation showed a complex and almost completely organized morphology.

Recent findings indicate that interleukin-1 beta (IL1 beta), a monokine secreted by stimulated macrophages and monocytes, modulates neuroendocrine functions in a manner similar to classical hormones. In this study we show that IL1 modulates PRL secretion, assessed by reverse hemolytic plaque assay, and describe the effect of the monokine on adenylate cyclase activity and calcium fluxes in rat normal pituitary cells. In basal and vasoactive intestinal peptide (VIP)-stimulated conditions, low doses of IL1 reduced the mean plaque area, a direct index of PRL secretion without affecting the percentage of PRL-secreting cells. Similarly, low concentrations of IL1 inhibited adenylate cyclase activity in both basal and VIP-stimulated conditions, while higher concentrations restored the enzymatic activity to the control value. IL1 also caused a biphasic effect on the free intracellular calcium increase induced by maitotoxin, a calcium channel activator, being inhibitory at low and stimulatory at high concentrations. The effects of IL1 on adenylate cyclase activity and calcium fluxes were reversed by preincubation of the monokine with its polyclonal antibody, thus confirming the specificity of the effects. In conclusion, our data show that IL1 modulates PRL secretion by acting directly on pituitary cells through interaction with the adenylate cyclase-cAMP system and calcium flux.

Tetranychus urticae is a phytophagous, macroscopic mite ubiquitous to warm climates, which causes occupational allergic symptoms in farm workers.This study was designed to investigate the cause and mechanisms of an otherwise "undetermined" recurrent and occupational dermatitis on the hands of farm workers and to obtain a better diagnostic profile of the urticaria that many farm workers who are allergic to T. urticae experience.Fifty-six farm workers with occupational cutaneous and/or respiratory symptoms and 10 healthy farm workers underwent in vivo and in vitro diagnostic trials by skin prick testing, serum-specific IgE determination, and three different types of epicutaneous tests.Four of the 10 patients with recurrent dermatitis had a positive IgE reaction to T. urticae, and five of the 10 had a positive patch test response. The percentage of positive patch test responses between subjects with positive T. urticae IgE and those with negative T. urticae IgE was similar. The diagnostic trial allowed the diagnosis of immune-mediated contact dermatitis in seven of the 10 patients with recurrent occupational dermatitis on the hands. The result of the epicutaneous open test on healthy skin with no history of eczema was positive in almost all patients with IgE to T. urticae and episodes of occupational urticaria.Urticaria frequently occurring in farm workers allergic to T. urticae is an allergic contact urticaria. T. urticae is also directly responsible for recurrent dermatitis, caused by a different immune-mediated mechanism, in farm workers.

The morpho-functional and topographical features of the lingual papillae situated on the dorsal surface of the bovine tongue, were studied utilizing LM and SEM techniques. In the bovine species, the functional differentiation of the lingual papillae in their gustatory and mechanical modes, seems to be related to the position of the papillae rather than to their morphological features. The gustatory function predominates over the mechanical one on the caudal tract of the tongue body (lateral to the lingual torus and associated with circumvallate papillae). The gustatory function also involves the conical papillae situated on the caudal tract of the tongue body. This gustatory function is aided by additional anatomical structures: 1. The grooves situated on the rostro-lateral side of the conical papillae; 2. The microcraters located on the top of the fungiform papillae; and 3. The furrows of the circumvallatae papillae vallum. The contact between saliva soluble food particles and taste buds is aided and made more efficient by these structures. Furthermore, a large variety of conical papillae were observed, whereas no filiform papillae were noted.

The Morpho-structural characteristics of the vallate papillae of the bovine tongue were studied utilizing LM, SEM and TEM techniques. In the bovine, the vallate papillae are numerous and different in size. They have a prominent vallum papillae which surrounds a deep sulcus papillae. The sulcus has many elongated taste buds on its inner wall. Nervous fibers enter the taste bud and sometimes have rosary shaped axons. The gustative sensitivity of these animals may be instrumental, in their choice of food and avoidance of noxious plants.

The goat choroid plexus angioarchitecture of the lateral ventricles was studied under the SEM using the method of "microvascular corrosion casts". The whole plexus is semilunar shaped and directed in an antero-posterior, latero-median fashion. In the plexus the lateral extremity is larger than the median one. All the components of the vascular bed (arteries, veins and capillaries) of the choroid plexus have interesting morpho-structural features. In particular, the capillaries are more developed than the other components and they are variously located on both sides of the plexus. The capillary network has a various organization in different zones of each side of the plexus.

The morphostructural characteristics of lingual papillae in 30 adult buffaloes were studied by SEM. In this species, dorsal lingual surface papillae shows different shape so that their classification is complicated. Topography and morphology of conical and vallate papillae presents significant aspects. Particularly, the conical papillae have a different morphological organization when they are localized over the side of the tongue apex or over dorso-lateral tract of the tongue body. The functional aspects are discussed.

It has been demonstrated that: a) part of the inhalant allergenic particles we normally breath, adhere to the oropharyngeal mucosa, and eventually progress to the gastrointestinal tract; b) digestive tract mucosa is able to produce specific IgE against aeroallergens even before than respiratory tract mucosa. The case is described of a 5-year-old girl who presented a daily vomiting since she was 6 months. All clinical instrumental and laboratory findings had been unable to reach a definite diagnosis. SPT (inhalants and foods): Dermatoph. pteronyssinus: + (confirmed by RAST). The patient had an immediate, complete recover just following the clinician's instruction for HDM domestic prevention. Symptoms appeared again in response to a NPT performed with Dermatophagoides extract. The positivity of the exclusion-re-exposure test confirmed the diagnosis of HDM-induced gastrointestinal allergic syndrome, so far not described in literature (to my knowledge). Immunological considerations: since it is known that patients allergic to HDM do not usually present a specific IgE-mediated gastrointestinal allergic syndrome, it is suspectable that an immunological tolerance can be instaured toward inhalant allergens as it normally happens toward food allergens. In atopic individuals there is a high expression of ICAM-1, VCAM-1 and other adhesion molecules on the surface of HEV at BALT level. Adhesion molecules expression and immunocompetent cells activation are modulated by several mechanisms among which the cytokine network plays a major role. The author speculates that sensitized lymphocytes may migrate from intestinal to bronchial mucosa, via lymphocytic immunoallergic competence. In the described clinical case this mechanism did not work.

The innervation of the Syrinx was studied in 15 female Peking Ducks of 10 to 12 months old by using L.M. and E.M. In this organ a nervous plexiform net is present located in the connective below the epithelium along the membrana tympaniformis medialis. This net-work is made of the nervous trunks which repeatedly anastomise among themselves and display during their course acapsulate ganglions. However, no specialized contact device were encountered at the periphery.

The microvasculature of the lingual mucosa of the goat was studied by SEM using microvascular corrosion casts. A complex microcirculatory system was observed to supply the large number of papillae, which had a wide structural variety. Generally, the organization of the vascular network in the lingual papillae was found to be influenced by the nature of the microcirculation in the lamina propria. Another characteristic feature noted was the presence of numerous valves in the venules situated in the lamina propria. They were oriented toward the broader side of the venules and probably control the rapid outflow of blood. This in turn most probably leads to an increased mucosal blood supply, which might sustain both mechanical and gustatory functions.

Neural receptors of the porcine clitoris were examined using light and electron microscopy. Perfusion with ink allowed study of the unique vascular arrangement associated with the genital corpuscles. Sensory nerve-endings were generally rounded and formed a morphologically and structurally characteristic unit. They were composed of a network of primarily non-myelinated nerve fibers and flat cells. Between these structures, isolated small blood vessels were embedded amongst collagen fibers and amorphous material. An external capsule of variable thickness always surrounded the structures. A complex arrangement of vessels in the center of the capsule was formed by a subpapillary mesh. The same arrangement was seen in the morphologically comparable penile genital corpuscles of the same species. Based on the particular arrangement of blood vessels and nerve fibers in the genital corpuscles, some authors postulate the presence of a neuro-vascular glomerulus.

The structural organization of the epithelium lining the clavicular air sac in Duck was studied by light and electron microscopic. This epithelium consists of two types of pavement like cells. The first flat types of cells are numerous with large mitochondria containing cytoplasm and elongated nuclei. The second short and dumpy cells are few in number and are after seen near the ostium. They could have long microvilli like pseudopod on their apical surface ready to engulf inhaled foreign particles. Desmosomal bridges too are found between contiguous cells.

The morphological and structural features of the vascular component of the bursa of Fabricius in the duck are described. By means of an intravasal perfusion of suitably colored neoprene Latex or Microscopaque, or a perfusion of Indian ink, it was possible to follow visually the routes of arteries, veins and the microcirculation; this last represents a major characteristic of the bursal follicle in the duck. The microcirculation has its origins in several roots arising from the follicular arterioles, which in the region of the cortex constitute an open vascular layer. From these pre-capillary arterioles, adjacent to the limiting layer between cortex and medulla, a dense capillary network is formed. With the aid of an electron microscope the presence of a cortico-medullary barrier of an epithelio-reticular nature was revealed; close to it there are the terminal branches of the intrafollicular network. The barrier appears to be structurally more compact than that of the cortical capillaries of the mammalian thymus.

The arrangement and the blood of Thymus lobes were shown by intra-vasal injection of Indian ink or Neoprene Latex. The origin and the distribution of the arteries were studied within thymic lobe and until the final branches represented by endolobulare capillaries. These branches represent the origin of the vein of the thymic lobe.

In the present study we evaluated the effect of interleukin 1-beta on adenylate cyclase activity in 235-1 pituitary cell line. The dose-response curve of interleukin 1 beta effect on adenylate cyclase activity showed a significant inhibition of basal enzyme activity at 1 pM concentration, while the inhibition of forskolin stimulated adenylate cyclase activity was more pronounced and evident at both 0.01 and 1 pM concentrations. The action of the monokine on basal enzyme activity was almost completely reverted by polyclonal anti interleukin 1 beta antibody. The incubation of the cells for 48h with interleukin 1 beta showed a different pattern of response. The inhibitory effect of interleukin 1 beta on adenylate cyclase activity disappeared, while the highest concentration of interleukin 1 beta tested, caused a meaningful stimulation of adenylate cyclase activity which is not present in acute condition. These data show that interleukin 1-beta interacts with the cAMP-generating system in the 235-1 clonal pituitary cells.

In the present paper we report that the ROHA -9 cell line, an Epstein-Barr virus (EBV)-transformed human B cell line with accessory cell capabilities, constitutively secretes a soluble factor with the biochemical and biological characteristics of human monocyte-derived IL-1. The IL-1 derived from ROHA -9 augmented murine thymocyte proliferation and enhanced the proliferative response of human T lymphocytes to concanavalin A (Con A). The ROHA -9-derived IL-1 activity eluted from Sephacryl S-200 in two peaks, at 15- 18K and 32- 35K mol wt, eluted from DEAE-Sephacel at 50-80 and 110-130 mM NaCl, and showed charge heterogeneity with peaks at pI 7.3, 6.1, and 4.1 on isoelectrofocusing (IEF). These findings suggest that B cells may elaborate an IL-1-like activity. During the logarithmic growth of ROHA -9 cells, a inhibitory factor that inhibited the response of mouse thymocytes to IL-1 was also produced. This factor had a mol wt of 95K on Sephacryl S-200, eluted at 150 mM NaCl on DEAE-Sephacel and showed a peak of pI 4.7 on preparative IEF. The inhibitory factor appeared to be selective in its effects on IL-1 responses, since it did not inhibit the activity of IL-2 on mouse thymocytes or on the growth of the IL-2-dependent CT6 cell line. This "contra-IL-1" inhibited the response of murine thymocytes to suboptimal (1 microgram/ml) but not optimal (10 micrograms/ml) doses of Con A and the response of human peripheral blood lymphocytes to streptolysin O ( SLO ) or to alloantigens. Moreover, the factor could be absorbed by mouse thymocytes but not by CT6 cells, and such thymocytes pretreated with contra-IL-1 failed to response to IL-1. Although this inhibitor is the product of a transformed B cell line, it may be representative of regulatory substances that normally control IL-1 activities either at the extracellular or intracellular level.

In the present study we investigated the effect of the interleukin 1 beta on intracellular free calcium concentrations in 235-1 cell line both in basal conditions and after stimulation by the calcium channel activator maitotoxin. Interleukin 1 beta (from 0.01 pM to 10 nM) was unable to significantly affect basal cytosolic free calcium levels in acute conditions. The preincubation of these cells with interleukin 1 beta for 48h modulates maitotoxin stimulation of calcium fluxes without modifying basal intracellular free calcium levels. Low concentrations of interleukin 1 beta (0.01 pM, 1 pM) caused a marked reduction of intracellular free calcium concentrations increase induced by maitotoxin while higher doses of the monokine potentiated maitotoxin stimulation of calcium fluxes. The specificity of interleukin 1 beta effect was tested by means of polyclonal anti-interleukin 1 beta antibody (titer 1:100) which significantly abolished the inhibitory effect of interleukin 1 beta on free cytosolic calcium levels. These results show that a long lasting interaction of interleukin 1 beta with its receptor is able to influence voltage-sensitive calcium channels activation induced by maitotoxin in 235-1 cells.

We studied the role of stimulant processing and presentation and of IL 1 in monocyte-mediated activation of human lymphoproliferative responses. The effects of two lysosomotropic agents, ammonium chloride and chloroquine, on the capacity of human monocytes to activate T lymphocyte responses to the soluble antigen streptolysin O (SLO) and to the polyclonal stimulant S. aureus protein A (SpA) were investigated. These agents inhibited the presentation of SLO and SpA by human monocytes in a dose-dependent manner. The inhibition occurred if monocytes were treated with ammonium chloride and chloroquine for 1.5 hr, starting only 30 min after exposure to the stimulants, whereas only minimal inhibition occurred when monocytes were treated with the two lysosomotropic compounds 2 hr after pulsing with SLO or SpA. In contrast, cell membrane alloantigen presentation by monocytes in the MLR was not affected by ammonium chloride or chloroquine treatment. Thus, these reversible inhibitors of monocyte phagosome-lysosome functions presumably interfere with intracellular processing of the stimulants but do not seem to interfere with alloantigen presentation at the cell surface. Furthermore, we investigated whether gently fixed monocytes were still capable of passively presenting stimulant or whether active metabolic processes as well as IL 1 were required. We observed that only monocytes treated with paraformaldehyde after SLO or SpA pulsing stimulated a proliferative response by T lymphocytes, provided 50 U/ml of partially purified human IL 1 were added back to cultures. In contrast, monocytes fixed before exposure to SLO or SpA were not able to stimulate T lymphocytes even if supplemented by IL 1. Taken together these data suggest that a finite incubation period is required for human monocytes to become able to present SLO or SpA to T lymphocytes. During this time the soluble stimulants presumably undergo some metabolic process in viable macrophages perhaps at the phagosome-lysosome level, to become recognizable by T lymphocytes.

The morphostructural features of the lymphatic circle of swine visceral pleura were investigated by means of vital dye injection in the pleural cavity. In the animals treated as mentioned previously it was possible to observe that in the visceral pleura the superficial lymphatic network had formed the very thick mesh, while the underlying pulmonary parenchyma was not interested. This behaviour lets us suppose that the drainage of the visceral pleura is mainly a direct one.

In this study we report that the ROHA-9 cell line, an IL 1-secreting EBV-transformed human B cell line, exhibits an autocrine pathway of growth. In fact, ROHA-9 cells spontaneously secreted an autoregulatory growth factor that co-purified with the constitutively secreted IL 1-like molecules. Accordingly, monocyte-derived human IL 1, free of other known biological activities, also stimulated the growth of ROHA-9 cells in a dose-dependent way. Human recombinant interleukin 2, recombinant IFN-alpha or IFN-gamma and purified IFN-beta were ineffective when used at concentrations up to 1 X 10(3) U/ml. Furthermore, mouse recombinant IL 1, HPLC-purified multi-colony stimulating factor and partially purified preparations of BCGF were ineffective when assayed for growth-promoting activity on ROHA-9 cells. Moreover, a rabbit polyclonal antibody and a mouse monoclonal antibody to human IL 1 molecules blocked the growth of ROHA-9 cells induced by the autologous growth factor and by human IL 1. Lastly, purified human IL 1 increased the clonal efficiency of ROHA-9 cells seeded at a low cell concentration, allowing the isolation of the ROHA-9MC3 subclone, which showed similar growth response specificity and was particularly sensitive to the mitogenic activity of human IL 1.

Interleukin 1 (IL 1) production by normal human B lymphocytes was investigated. Normal human peripheral blood B lymphocytes were purified by sequential separation with the use of Ficoll-Hypaque gradient centrifugation, sheep red blood cell rosette formation, Percoll gradients, and treatment with monoclonal antibodies (anti-Leu-M1, B73.1, and T101) and complement. Both purified large B lymphocytes (BL) and small B lymphocytes (BS) produced IL 1-like (thymocyte co-mitogenic and fibroblast mitogenic) activities in response to lipopolysaccharide. Maximal production of IL 1 activity by both BL and BS occurred at 48 hr. The m.w. of IL 1 activities from both BL and BS were about 20,000 with high pressure liquid chromatography, and the major isoelectric point of BL- and BS-derived IL 1 activity was 7.0. A rabbit anti-human monocyte IL 1 antiserum inhibited the activity of B cell-derived IL 1, suggesting antigenic similarities of monocyte- and B lymphocyte-derived IL 1 moieties. These data suggest that normal B lymphocyte-derived IL 1 activity is biochemically and immunologically similar to monocyte-derived IL 1.

The innervation of the mucous membranes in the paranasal cavities of "ovis aries" appears homogeneous as regards the various nasal sinuses. It is represented by numerous nervous bundles which are in the deep and middle parts of the mucous membranes, and have sensory terminals and fibres assigned to the vessels and glandular adenomeres. In the subepithelial stratum can be noted a superficial plexus from which come off the fibres assigned to the epithelial elements of the mucous membranes.

The microcirculation of thymic lobule has been studied. The vascular feature is markedly different compared with mammalian thymic circulation. Only one artery trunk, the intralobular thymic artery branching off from intralobular artery has been evidentiated. Interlobular arter penetrates the lobule and originates a bed of narrow thin-walled capilary network, thinner in the pheriphery.

The structural and histological characteristics of the limiting layer between the cortex and medulla of the bursa of Fabricius in the duck are described in relation to different functional moments of the organ. On E.M. study the limiting layer, in duck having 30 days of age, is constituted of reticular-epithelial cells palisades in shape with many cytoplasmics extension. Lymphatic migration occur in interstitial spaces.

The lymphatic component in the pharynx of Buffalo presents some peculiar characteristics of species above all as regards the palatine tonsil. This is characterized by the presence of numerous lobes, by the fast pigmentation of the mucosa in the sinus and sometimes by the crypts and by the wealth of glandular adenomeres. The lingual tonsil, the tonsil of the soft palate, the pharyngeal tonsil and the tubal tonsil are represented by lymphatic elements arranged in the lamina propria of the mucosa and only sometimes they are organized as follicles.

Neoplastic cells obtained from the pleural effusion of a patient with Hodgkin's disease have been maintained in culture since 1978. These tumor cells have been shown to have the cytologic features, cytochemical staining, and cell surface markers of Reed-Sternberg cells. In this study we demonstrate that the cell line termed L428 is a potent stimulator of the primary human mixed lymphocyte reaction. Significant proliferation occurred when mononuclear leukocytes obtained from normal donors were stimulated with radiated L428 cells at responder:stimulator ratios varying from 200:1 to 20:1. Proliferative responses occurred between days 3 and 6 of the cultures with maximal proliferation on day 5. Under optimal culture conditions, mean net proliferative response of 14 normal donors was 51,000 +/- 10,600 dpm. The mixed lymphocyte response was totally blocked by concentrations of monoclonal anti-Ia antibody that had no effect on concanavalin A-induced proliferation. However, the mixed lymphocyte response was not blocked by an anti-K562 cell monoclonal antibody of the same immunoglobulin subclass that binds to the L428 cells. Antigen processing by responder monocytes or Ia-positive cells was not required for the MLC. When responder T cells from two normals were depleted of Ia-bearing cells and monocytes, the mixed lymphocyte reaction between the two normals was eliminated, yet the stimulation of each normal by the L428 cells was not reduced. The cells that proliferated in response to stimulation by the L428 cells were T cells, primarily of the helper subset. No IL 1 activity could be detected in concentrated supernatants of L428 cultures after stimulation of L428 cells by mitogens, phorbol esters, or muramyl dipeptide, or in the MLC. All of these cultures contain fetal calf serum. However, the L428 cells are capable of producing IL 1, because IL 1 was detected when the L428 cells were stimulated with LPS in the absence of fetal calf serum. These neoplastic cells, obtained from Hodgkin's disease, have many similarities to the murine as well as human dendritic cells.

In bovine bronchia a peribronochial nervous plexus, two thin one around polmonary arteries and veins and an inner plexus disposed in the mucous structures have been evidentiated. Numerous big nervous ganglia in connection with peribronchial plexus, particularly in the wider bronchia have been observed: in the mucous and submucous membrane of wider bronchia, sensitive nervous terminations are located.

The Bridging Sheet domain of HIV-1 gp120 is highly conserved among the HIV-1 strains and allows HIV-1 binding to host cells via the HIV-1 coreceptors. Further, the bridging sheet domain is a major target to neutralize HIV-1 infection. We rationally designed four linear peptide epitopes that mimic the three-dimensional structure of bridging sheet by using molecular modeling. Chemically synthesized peptides BS3 and BS4 showed a fair degree of antigenicity when tested in ELISA with IgG purified from HIV+ broadly neutralizing sera while the production of synthetic peptides BS1 and BS2 failed due to their high degree of hydrophobicity. To overcome this limitation, we linked all four BS peptides to the COOH-terminus of GST protein to test both their antigenicity and immunogenicity. Only the BS1 peptide showed good antigenicity, however, no envelope specific antibodies were elicited upon mice immunization. Therefore we performed further analyses by linking BS1 peptide to the NH2-terminus of the E2 scaffold from the Geobacillus Stearothermophylus PDH complex. The E2-BS1 fusion peptide showed good antigenic results, however only one immunized rabbit elicited good antibody titers towards both the monomeric and oligomeric viral envelope glycoprotein (Env). In addition, moderate neutralizing antibodies response was elicited against two HIV-1 clade B and one clade C primary isolates. These preliminary data validate the peptide mimotope approach as a promising tool to obtain an effective HIV-1 vaccine.

The inhibitor of Bruton tyrosine kinase γ (IBtkγ) is a negative regulator of the Bruton tyrosine kinase (Btk), which plays a major role in B-cell differentiation; however, the mechanisms of IBtkγ-mediated regulation of Btk are unknown. Here we report that B-cell receptor (BCR) triggering caused serine-phosphorylation of IBtkγ at protein kinase C consensus sites and dissociation from Btk. By liquid chromatography and mass-mass spectrometry and functional analysis, we identified IBtkγ-S87 and -S90 as the critical amino acid residues that regulate the IBtkγ binding affinity to Btk. Consistently, the mutants IBtkγ carrying S87A and S90A mutations bound constitutively to Btk and down-regulated Ca2+ fluxes and NF-κB activation on BCR triggering. Accordingly, spleen B cells from Ibtkγ−/− mice showed an increased activation of Btk, as evaluated by Y551-phosphorylation and sustained Ca2+ mobilization on BCR engagement. These findings identify a novel pathway of Btk regulation via protein kinase C phosphorylation of IBtkγ.