Immunophenotypic analysis of human spleen compartments.

Microanatomical compartments of the human spleen are yet under evaluation as most of the present information comes from experiments on animals with different anatomical structures. Immune staining of stromal and blood-born cells by cell surface antigens facilitates the differentiation of functional microanatomical compartmentalization of immune organs, including the spleen. Twenty-two specimens from healthy adult subjects with the average age of 35.6 +/- 13.8 (Range 17 to 58) years were included in this study. Monoclonal antibodies used in this study were supplied from the 5th, 6th and 7th International Workshops and Conferences on Human Leukocyte Differentiation Antigens. Tetraspan antigens presented a rather unique staining pattern in the human spleen, suggesting special roles for each (CD9, CD53, CD63, CD151 and CD231) in certain locations. Sinus lining cells presented a distinctive antigenic profile, sharing both endothelial cell (CD31, CD36, CD54, CD62P, CD102, CD105, CD106 and CD146) and macrophage lineage characteristics. The sheathed capillaries were not restricted to the perifollicular zone alone. Extracellular matrix receptors (CD49 a, CD49 b, CD49 c, CD49 e, CD49f, CD29 and CD44) stained the penicillary arterioles and vascular smooth muscle. These molecules were also found on the vascular endothelium. Leukocyte antigens (CD11a, CD11b, CD22, CD43, CD45, CD45RB, CD45RO and CD50) were mainly expressed in the white and red pulp of the spleen at different intensities, excluding the penicillary arterioles. Activation antigens (CD26, CD71 and CD98) presented a diffuse and broad staining pattern. In conclusion, microanatomical compartmentalization, microcirculation and function of the human spleen were evaluated using a wide panel of monoclonal antibodies.     

Functional uncoupling of hemodynamic from neuronal response by inhibition of neuronal nitric oxide synthase.

The cerebrovascular coupling under neuronal nitric oxide synthase (nNOS) inhibition was investigated in alpha-chloralose anesthetized rats. Cerebral blood flow (CBF), cerebral blood volume (CBV), and blood oxygenation level dependent (BOLD) responses to electrical stimulation of the forepaw were measured before and after an intraperitoneal bolus of 7-nitroindazole (7-NI), an in vivo inhibitor of the neuronal isoform of nitric oxide synthase. Neuronal activity was measured by recording somatosensory-evoked potentials (SEPs) via intracranial electrodes. 7-Nitroindazole produced a significant attenuation of the activation-elicited CBF (P<10(-6)), CBV (P<10(-6)), and BOLD responses (P<10(-6)), without affecting the baseline perfusion level. The average DeltaCBF was nulled, while DeltaBOLD and DeltaCBV decreased to approximately 30% of their respective amplitudes before 7-NI administration. The average SEP amplitude decreased (P<10(-5)) to approximately 60% of its pretreatment value. These data describe a pharmacologically induced uncoupling between neuronal and hemodynamic responses to functional activation, and provide further support for the critical role of neuronally produced NO in the cerebrovascular coupling.     

Effects of probucol in hyperlipidemic rabbit liver: a preliminary ultrastructural study

Probucol is a lipid-lowering agent with an antioxidant effect; however, its influence on the liver remains unclear. The effects of probucol on hyperlipidemic rabbit liver are investigated to add a structural data on its therapeutical profile. Local albino rabbits were divided into three groups. 1) Hyperlipidemic group: fed with 1% cholesterol (150 g/kg/day) enriched chow for 2 months. 2) Probucol treated group: group 1 + intraperitoneal probucol (10 mg/kg/day) administration for 15 days. 3) Control group fed with normal chow. The blood lipid profile was investigated biochemically. Liver samples were examined electronmicroscopically. Within the parenchymal cells of group 1, the amount of rough surfaced endoplasmic reticulum was increased, its cisterna was dilated displaying a moderately electron dense substance in it and showed close apposition with the condensed mitochondria. In group 2, smooth surfaced endoplasmic reticulum was in extensive amounts filling almost all of the cytoplasm, displayed a reticular, degenerated appearance and was in close relation with the condensed, degenerated mitochondria. Probucol may cause degenerative changes on the liver parenchyme at the subcellular level. It alters the structure of these cells mainly acting on the smooth surfaced endoplasmic reticulum and the mitochondria that are known to be involved in cellular detoxification.     

Collagen-chondroitin sulfate-based PLLA-SAIB-coated rhBMP-2 delivery system for bone repair

Bone morphogenetic proteins (BMPs) are osteoinductive proteins used intensively in clinical investigations involving various bone-related treatments. Owing to their high potential in new bone formation they require local application at the treatment site. For this purpose various controlled delivery systems with BMPs as the excipients have been prepared in recent years. Focusing on this clinical need a disc-shaped BMP carrier was designed as a local delivery system using soluble collagen and chondroitin sulfate. In situ release studies carried out with a model protein (FITC-labeled Protein A) presented a very high rate of release; with most of the protein content being released within 24 h. This rate could be decreased by providing a poly(L-lactide) (PLLA) and sucrose acetate isobutyrate-based (SAIB-based) coat around the release system, applied after BMP loading. In this way, it was possible to extend the release period from 24 h to about 12 days. In situ release of BMP from the same carriers, as quantitated using an ELISA kit, was even slower, with 50% of the protein being released in 15 days. In order to be able to secure the BMP delivery system at the bone defect site and to provide support a mesh knitted using Vicryl sutures and bonded with poly(L-lactide-co-glycolide) (PLGA) was tested in in vivo. Two time periods, 1 and 3 weeks, were used to evaluate the healing process. Osteoinduction by the BMP carrier system was assessed by histology-based bone scoring and X-ray examinations. PLLA-SAIB-coated collagen discs containing BMP presented good biocompatibility and optimum osteogenic stimulation. Structural changes in histological micrographs at week 1 indicated dose-dependent periosteal ossification. At the end of week 3 histological findings with both BMP (1 and 2 microg) doses were almost the same.     

Inactivation of a turkey herpesvirus by ultra-violet light

The sensitivity to inactivation by ultraviolet light of the PB 1 strain of herpesvirus of turkeys (HVT) was determined and compared with the relative sensitivity of pseudo-rabies virus (PRV), another member of the herpesvirus group. HVT (PB1) was 6.3 times more resistant than PRV when assayed in chick embryo fibroblasts. The survival curve of HVT (strain PB1) had one component and the survival curve of PRV had two components. Survival curves of HVT and PRV in chick embryo fibroblasts treated with caffeine and in duck embryo fibroblasts are also presented. Evidence was obtained for host cell reactivation of HVT and PRV in chick embryo fibroblasts. Multiplicity reactivation could be clearly demonstrated for UV-irradiated PRV, but not for HVT.     

Hepatoprotective Effect of Mixture of Dipropyl Polysulfides in Concanavalin A-Induced Hepatitis

The main biologically active components of plants belonging to the genus Allium, responsible for their biological activities, including anti-inflammatory, antioxidant and immunomodulatory, are organosulfur compounds. The aim of this study was to synthetize the mixture of dipropyl polysulfides (DPPS) and to test their biological activity in acute hepatitis. C57BL/6 mice were administered orally with DPPS 6 h before intravenous injection of Concanavalin A (ConA). Liver inflammation, necrosis and hepatocytes apoptosis were determined by histological analyses. Cytokines in liver tissue were determined by ELISA, expression of adhesive molecules and enzymes by RT PCR, while liver mononuclear cells were analyzed by flow cytometry. DPPS pretreatment significantly attenuated liver inflammation and injury, as evidenced by biochemical and histopathological observations. In DPPS-pretreated mice, messenger RNA levels of adhesion molecules and NADPH oxidase complex were significantly reduced, while the expression of SOD enzymes was enhanced. DPPS pretreatment decreased protein level of inflammatory cytokines and increased percentage of T regulatory cells in the livers of ConA mice. DPPS showed hepatoprotective effects in ConA-induced hepatitis, characterized by attenuation of inflammation and affection of Th17/Treg balance in favor of T regulatory cells and implicating potential therapeutic usage of DPPS mixture in inflammatory liver diseases.     

Antioxidative vitamin treatment: Effect on lipid peroxidation and limb swelling after revascularization operations

The objective of this study was to evaluate the antioxidative properties of the multivitamin cocktail Omnibionta^® (-tocopherol, ascorbic acid, retinol, vitamin B complex) in terms of diminishing lipid peroxidation with improvement of leg edema performance after limb revascularization operations in humans. Fifty-one subjects were selected; the control group contained 27 patients and the treatment group 24 patients, who received the vitamin cocktail intravenously before the start of reperfusion. All patients suffered from acute or chronic arterial occlusive disease, except two subjects with arterial trauma. MDA-TBARS in plasma, quantified by HPLC, taken as a measure of lipid peroxidation was significantly increased (p<0.001) in the control group 1 hour after reperfusion onset and decreased to its baseline value within the following 2 hours (0.73±0.26, 1.21±0.48, 0.99±0.48, 0.73±0.33 nmol/ml). In contrast, in the treatment group MDA-TBARS did not exceed the baseline value during the reperfusion period (0.93±0.30, 0.70±0.29, 0.65±0.23, 0.70±0.37 nmol/ml). Leg edema, expressed by extremity circumference, was significantly (p<0.008) elevated in the control group (30.7±4.04 cm versus 35.35±4.12 cm) compared to a lack of increase in the treatment group (29.25±5.13 cm versus 29.76±5.70 cm). These results suggest that antioxidative vitamin treatment might be valuable in preventing lipid peroxidation and decreasing extremity edema.

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Resumen El objeto del presente estudio fue evaluar las propiedades antioxidantes del coctel multivitamínico Omnibionta^® que contiene las vitaminas -tocoferol, ácido ascórbico, retinol y el complejo B, en cuanto a disminuir la peroxidación lipídica y reducir el edema en las operaciones de revascularización de extremidades en humanos. Se seleccionaron 51 individuos: el grupo control de 27 pacientes y el grupo de tratamiento de 24 pacientes que recibieron el coctel multivitamínico por vía intravenosa antes del comienzo de la reperfusión. Todos los pacientes exhibían enfermedad arterial oclusiva aguda o crónica, excepto 2 pacientes con trauma arterial. Se hicieron determinaciones de MDA-TBARS en el plasma, cuantificado por HPLC, como medida de la peroxidación lipídica, encontrándose que se hallaba significativamente aumentada (p<0.001) en el grupo control 1 hora luego del comienzo de la reperfusión y que descendió hasta los niveles basales en el curso de las siguientes 2 horas (0.73±0.26, 1.21±0.48, 0.99±0.48, 0.73±0.33 nmol/ml). El edema de la pierna, según la circunferencia de las extremidades, apareció significativamente elevado (p<0.008) en el grupo control (30.7±4.04 vs. 35.35±4.12 cm) en comparación con el ningún aumento en el grupo de tratamiento (29.25±5.13 vs. 29.76±5.70 cm). Tales resultados sugieren que el tratamiento vitamínico antioxidante puede ser de valor en la prevención de la peroxidación lipídica con disminución del edema de la extremidad.

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Résumé L'objectif de cette étude a été d'évaluer les propriétés antioxydatives du cocktail multivitamine Omnibiota^®, contenant les vitamines -tocophérol, de l'acide ascorbique, du rétinol et du complexe vitamine B, pour diminuer les peroxydations lipidiques et améliorer l'oedème de jambe après reconstruction vasculaire chez l'homme. Cinquante et un sujets ont été sélectionnés, 27 formant le groupe témoin, et 24 patients recevant le cocktail en intraveineux avant la reconstruction vasculaire. Tous les patients avaient une maladie artérielle occlusive chronique, sauf deux qui souffraient d'un traumatisme artériel. Le MDA-TBARS dans le plasma, quantifié par HPLC, comme mesure de la peroxydation lipidique, était significativement augmenté (p<0.001) dans le groupe contrôle une heure après le début de la reperfusion et a diminué jusqu'aux valeurs du départ dans les deux heures suivantes (0.73±0.26, 1.21±0.48, 0.99±0.48, 0.73±0.33 nmol/ml). Dans le groupe traité, le MDA-TBARS n'a pas dépassé les valeurs de base pendant la période de reperfusion (0.93±0.30, 0.70±0.29, 0.65±0.23, 0.70±0.37 nmol/ml). L'oedème des membres inférieurs, exprimé en circonférence de l'extrémité de la jambe, était signficativement plus augmenté (p<0.008) dans le groupe contrôle (30.7±4.04 vs. 35.35±4.12 cm) que dans le groupe traité (29.25±5.13 vs 29.76±5.70 cm). Ces résultats suggèrent que le traitement vitaminique antioxydatif pourrait être utile dans la prévention de la peroxydation lipidique, réduisant ainsi l'oedème des extrémités.

     

Pentadecapeptide BPC 157 attenuates disturbances induced by neuroleptics: the effect on catalepsy and gastric ulcers in mice and rats.

A gastric pentadecapeptide, BPC 157, with the amino acid sequence, Gly-Glu-Pro-Pro-Pro-Gly-Lys-Pro-Ala-Asp-Asp-Ala-Gly-Leu-Val, MW 1419, known to have a variety of protective effects in gastrointestinal tract and other organs, was recently shown to particularly affect dopamine systems. For instance, it blocks the stereotypy produced acutely by amphetamine in rats, and the development of haloperidol-induced supersensitivity to amphetamine in mice. Consequently, whether pentadecapeptide BPC 157, that by itself has no cataleptogenic effect in normal animals, may attenuate the immediate effects of neuroleptics application, particularly catalepsy, was the focus of the present report. Prominent catalepsy, otherwise consistently seen in the mice treated with haloperidol (0.625, 1.25, 2.5, 5.0 and 10.0 mg/kg b.w., i.p.) and fluphenazine (0.3125, 0.625, 1.25, 2.5 and 5.0 mg/kg b.w., i.p.) after 1.5, 3, 4.5, 6 and 7.5 h following administration, was markedly attenuated when pentadecapeptide BPC 157 (10 microg or 10 ng/kg b.w., i.p.) was coadministered with the neuroleptic. The number of cataleptic mice was markedly lower throughout most of the experimental period. Moreover, on challenge with lower doses of neuroleptics, catalepsy appearance was postponed and the mice, otherwise cataleptic since the earliest period, became cataleptic later, not before 3 or 4.5 h after neuroleptic administration, especially if protected with higher pentadecapeptide dose. Besides catalepsy, coadministration of the pentadecapeptide BPC 157, given in the above mentioned doses, reduced not only catalepsy but somatosensory disorientation (for 7.5 h after administration of a neuroleptic, assessed at intervals of 1.5 h, by a simple scoring system [0-5]) in haloperidol- or fluphenazine-challenged mice as it did in mice treated with sulpiride (20, 40, 80 and 160 mg/kg b.w., i.p.) or with clozapine (25, 50 and 100 mg/kg b.w., i.p.), in which case catalepsy was absent. In other experiments, considering the gastric origin of this pentadecapeptide, the focus was shifted to the evidence that a dose of haloperidol, cataleptogenic due to dopamine receptors blockade, induces gastric ulcers in rats. Coadministration of pentadecapeptide BPC 157 (10 microg, 10 ng, 1.0 ng, 100 pg/kg b.w., i.p.) to rats completely inhibited the lesions otherwise regularly evident 24 h after haloperidol (5.0 mg/kg b.w., i.p.) in control rats (18 of 20 rats had gastric lesions). This activity accompanied the antagonism of the haloperidol catalepsy in rats (assessed at 60-min intervals from I to 5 h after haloperidol), when 10-microg- or 10-ng regimens were given (lower doses could not influence catalepsy). Together, these findings indicate that pentadecapeptide BPC 157 fully interacts with the dopamine system, both centrally and peripherally, or at least, that BPC 157 interferes with some steps involved in catalepsy and/or ulcer formation.