Cinaciguat prevents neointima formation after arterial injury by decreasing vascular smooth muscle cell migration and proliferation.

AIMS: Vascular smooth muscle cell (VSMC) migration, proliferation and remodeling of the extracellular matrix contribute to lumen loss after arterial injury leading to restenosis. Several studies indicated the role of the cyclic guanosine monophosphate signaling in neointimal formation. Cinaciguat, the novel soluble guanylate cyclase activator, currently being in phase IIb clinical trial, has been shown to exert antiplatelet and anti-remodeling effects in animal models of vascular pathology. In this study we investigated the effects of cinaciguat on post-injury arterial stenosis. METHODS AND RESULTS: Male Sprague-Dawley rats (n=100) underwent endothelial denudation by wire injury of the right common carotid artery. Cinaciguat (10mg/kg/day orally) were administered to 50 rats (1-, 2-, 3-day and 1-, 3-week treatment time), while 50 rats received placebo. A 3-week treatment resulted in a significantly reduced vascular stenosis (17.53 ± 10.84% in the treatment group vs. 43.25 ± 30.83% in the control wire injury group) and neointima/media area ratio (0.45 ± 0.32 in the treatment group vs. 1.09 ± 0.69 in the control wire injury group). By using quantitative real-time PCR, Western blot and immunohistochemistry, matrix-metallopreoteinase-9 (MMP-9) was found to be upregulated in the control-injured carotids over the whole follow-up, and cinaciguat significantly decreased MMP-9 expression by 3 weeks. As assessed by protein immunoblot, injury-induced local decrease of soluble guanylate cyclase ß1 subunit could be recovered by cinaciguat. In vitro wound healing assay with VSMCs revealed dose-dependent antimigratory and antiproliferative effects of cinaciguat. Plasma level of cyclic guanosine monophosphate was significantly elevated after 3 weeks of treatment. CONCLUSION: Our results show that cinaciguat prevents injury-induced neointimal hyperplasia by decreasing VSMC migration and proliferation through the regulation of MMP-9.

Experiences with a large-size WWTP based on activated sludge-biofiltration processes: 25 months of operation.

The South-Budapest Wastewater Treatment Plant (SBWWTP) had been operated as a high-load activated sludge (AS) plant since the middle of the 60s. According to the requirements proposed by the water authorities the treatment process had to be upgraded into nutrient (phosphorus and nitrogen) removal. The upgrade of the plant comprised implementation of BIOFOR type nitrifying (NP) and post-denitrifying (DN) biofilters downstream of the AS stage. Phosphorus removal was obtained by chemical precipitation that can be done at five different points for feeding ferric-sulfate (Fe2(SO4)3). Partial flow recirculation was administered from the nitrifying BIOFOR unit ahead of the AS basin for pre-denitrification utilizing raw wastewater as carbon source. The plant performance was monitored since the test operation period for 25 months. Experience revealed that significant nitrification occurs in the high-load activated sludge basin originally designed for carbon removal. During the summer period (characterized by temperature of 20-25 degrees C) about 37-42% ammonium conversion rate was observed in the reactor. The decreasing temperature in the wintertime resulted in lower nitrification rates, of about 6-10%. The combined activated sludge-biofiltration process proved its viability in the removal of organic matter, nitrogen and phosphorus. In this special configuration the AS system plays a key role in the nitrogen and organic matter removal.

The effect of leukocyte interleukin injection (Multikine) treatment on the peritumoral and intratumoral subpopulation of mononuclear cells and on tumor epithelia: a possible new approach to augmenting sensitivity to radiation therapy and chemotherapy in oral cancer--a multicenter phase I/II clinical Trial.

OBJECTIVES/HYPOTHESIS: The main objective of this study was to investigate the effect of the administration of a novel immunoadjuvant, leukocyte interleukin injection, as part of an immuno-augmenting treatment regimen on the peritumoral and intratumoral subpopulations of the tumor infiltrating mononuclear cells and on the epithelial and stromal components, when administered to patients with advanced primary oral squamous cell carcinoma classified as T2-3N0-2M0, as compared with disease-matched control patients (not treated with leukocyte interleukin injection). STUDY DESIGN: Multicenter Phase I/II clinical trial. Fifty-four patients from four clinical centers were included in the dose-escalating study (27 in each group [leukocyte interleukin injection-treated and control groups]). Cumulative leukocyte inter-leukin injection doses were 2400, 4800, and 8000 IU (as interleukin-2 equivalent). METHODS: Paraffin-embedded tumor samples obtained at surgical resection of the residual tumor (between days 21 and 28 after treatment initiation) were used. Histological analysis, necrosis evaluation, and American Joint Committee on Cancer grading were performed from H&E-stained sections. Immunohistochemical analysis was performed on three different tumor regions (surface, zone 1; center, zone 2; and tumor-stroma interface, zone 3). Trichrome staining was used to evaluate connective tissue, and morphometric measurements were made using ImagePro analysis software. Cell cycling was determined by the use of Ki-67 marker. RESULTS: Leukocyte interleukin injection treatment induced a shift from stromal infiltrating T cells toward intraepithelial T cells and posted a significant (P <.05) increase in intraepithelial CD3-positive T cells independent of the leukocyte interleukin injection dose, whereas the increase in CD25 (interleukin-2 receptor alpha [IL-2Ralpha])-positive lymphoid cells was significant only at the lowest leukocyte interleukin injection dose (P <.05). Furthermore, both low- and medium-dose leukocyte interleukin injection treatment induced a significant (P <.05) increase in the number of cycling tumor cells, as compared with control values. CONCLUSION: The results could be highly beneficial for patients with oral squamous cell carcinoma. First, leukocyte interleukin injection treatment induces T-cell migration into cancer nests and, second, noncycling cancer cells may enter cell cycling on administration of leukocyte interleukin injection. This latter effect may modulate the susceptibility of cancer cells to radiation therapy and chemotherapy. The findings may indicate a need to re-evaluate the way in which follow-up treatment (with radiation therapy and chemotherapy) of patients with head and neck cancer is currently approached.

Endogenous lipoid pneumonia associated with undifferentiated connective tissue disease (UCTD).

BACKGROUND: Lipoid pneumonia is a rare pulmonary disease, a form of pneumonia that has no classical radiological appearance, thus it can imitate other lung diseases. Lipoid pneumonia is usually classified into two major groups, depending on whether the source of oil/fat in the respiratory tract is from an exogenous or endogenous source. Undifferentiated connective tissue disease is a term used by rheumatologists to define a group of diffuse connective tissue disorders that lack definitive characteristics of any particular well-defined disorder. MATERIAL AND METHODS: A case study is reported of concomitant undifferentiated connective tissue disease and endogenous lipoid pneumonia. RESULTS: Histologically the macrophages appeared filled with lipid and were similar to atherosclerotic foam cell macrophages. Antibiotic and antimycotic treatments were ineffective. However, with concomitant steroid treatment, the patient exhibited absence of lung infiltration as well as other symptoms and was discharged. Therefore it is concluded that the lipoid pneumonia was steroid dependent. CONCLUSION: Since the patient's condition responded to steroid treatment, and it is clear that steroids inhibit phospholipase activity, the authors speculate that the subsequent decreased endoperoxide production may diminish lipid uptake by macrophages via decreasing modification of LDL or other lipid sources.

Effects of cocaine on the contents of neurohypophyseal hormones in the plasma and in different brain structures in rats.

The effects of acute and chronic cocaine treatments on the levels of the neurohypophyseal hormones oxytocin (OXT) and vasopressin (AVP) in the plasma and in different brain structures in rats were measured by radioimmunoassay (RIA). Acute cocaine treatment had no effect on the level of OXT in the plasma or in the amygdala, but increased OXT contents were measured in the hypothalamus and in the hippocampus. The OXT levels in the basal forebrain structures (including the septum and the nucleus accumbens) were decreased by a single dose of cocaine. The acute injection of cocaine increased the level of AVP in the plasma, and decreased contents of OXT were measured in the amygdala and in the basal forebrain. Repeated treatment with cocaine decreased the level of OXT in the plasma, hypothalamus and hippocampus. The AVP contents were decreased in all of the brain structures investigated, but no change was caused in the plasma level of AVP by repeated injections of cocaine. These results demonstrate complex, region-specific interactions between cocaine and the neurohypophyseal hormones in the brain and in the periphery underlying the alteration in behavioral and autonomic functions caused by acute and chronic cocaine exposure.

Chronic benzodiazepine administration. IX. Attenuation of alprazolam discontinuation effects by carbamazepine.

Clinical studies suggest that carbamazepine may attenuate effects of alprazolam discontinuation. Since discontinuation of chronic alprazolam in a mouse model is associated with behavioral alterations and upregulation at the gamma-aminobutyric acidA (GABAA) receptor, we studied the effects of carbamazepine administration after alprazolam (2 mg/kg/day) discontinuation. Open-field activity was increased in mice 4 days after alprazolam discontinuation, but this effect was reduced significantly by continuous infusion of carbamazepine, 25 or 100 mg/kg/day. Benzodiazepine receptor binding in vivo was increased in cortex at 2 and 4 days after alprazolam discontinuation, and in hypothalamus at 4 days; with carbamazepine, 100 mg/kg/day, binding in both regions at these time points was similar to control values. Similar results were observed in cortex with benzodiazepine receptor binding in vitro. GABA-dependent chloride uptake was also increased at 4 days alprazolam administration. Treatment with carbamazepine attenuated (P less than 0.10) this increase. Carbamazepine alone after vehicle did not alter benzodiazepine binding or GABA-dependent chloride uptake. These results indicate that carbamazepine administration after alprazolam discontinuation attenuates behavioral and neurochemical alterations associated with discontinuation.

Experimental stimulation of osteogenesis induced by bone matrix.

Bone gaps in 20 rabbits were filled with decalcified homologous bone. Direct currents, intravenous calcium, and anabolic steroids were administered for stimulation of osteogenesis. The shape and microscopic structure of the newly formed bone, together with the velocity of the osteogenesis, were investigated. Results were assessed by microscopic and radiographic analyses. In the group of calcium posttreatment, irregular shape and callosities were characteristic. Anabolic steroids failed to improve consolidation significantly; best results were seen after electrical stimulation of the grafted areas. In the group with electrical stimulation, fair shape, together with a more regular microscopic structure of the newly formed bone, were characteristic 4 weeks after surgery. Ungrafted gaps, together with defects filled with undecalcified cortical bone, failed to heal within the examined period of time.

The clinical use of aluminium oxide bioceramic implants for mandibular condyle replacement: a six year report.

In a 6-year period, six bilateral and five unilateral compact aluminium oxide ceramic condyle fixed with titanium screws were implanted. During the years following the operation, it was not necessary to remove any ceramic condyle. In only one case (rheumatic polyarthritis) was it necessary to reoperate on the patient because of bilateral reankylosis. Ceramics can be recommended in a wide field application because of the simplicity, uniformity, durability, and good tissue-enduring ability. The danger of postoperative ankylosis occurs only in patients suffering from rheumatoid arthritis.

Application of compact aluminum oxide ceramic implants in maxillofacial surgery.

In a 4-year period, 40 compact aluminum oxide ceramic implants fixed with titanium plates and screws were used: 24 for the correction of mandibular defects, 9 for temporomandibular joint replacement, and 7 for augmentation of facial contour. During the 1 to 4 years following the operations, it was not necessary to remove any aluminum oxide joint or implant used for contour correction. In 5 of the 24 cases of mandibular reconstruction, however, it was necessary to remove the ceramic. In 3 cases, this was due to recurrence of the tumor, and in 2 cases the cause was dermal necrosis.

Effects of beta-endorphin2-9 on arginine-8-vasopressin and oxytocin levels in hypothalamic and limbic brain regions.

Immunoreactive arginine-8-vasopressin (AVP) and oxytocin (OXT) were measured in rat hypothalamic and limbic brain regions after the intracerebroventricular administration of beta-endorphin fragment 2-9 (beta E2-9). The peptide decreased the AVP content of the hippocampus and the OXT levels in the septum and amygdala. The present data favor the view that beta E2-9 interacts with limbic AVP- and OXT-systems.

The effect on brain 5-HT of central lysine-vasopressin administration into different cerebral ventricular compartments depends on the site of injection.

The influence of lysine-8-vasopressin (LVP) on the steady-state level of serotonin was studied in different brain regions four hours after microinjection into the lateral ventricle (icv) or cisterna magna (cm). LVP (200 pg) increased the serotonin (5-HT) level in the hypothalamus and mesencephalon after cm application, while icv administration caused an opposite tendency. The findings suggest that the site of peptide administration is of decisive importance in the mediation of various effects of LVP on the transmitter metabolism. A possible link exists between opposite effects of cm versus icv injection of LVP on central serotoninergic mechanisms, as well as on behavioral and autonomic correlates of these treatment.

Elemental concentrations in human erythrocytes and blood plasma following radiotherapeutic irradiation.

Elemental concentrations in whole blood samples from irradiated mice were found [1] to change uncorrelated with the depression of red blood cell count. The aim of the present work was to clear up whether or not similar effects appear in the case of human radiotherapeutic irradiations. Concentration ratios were taken for erythrocyte and plasma fractions with the proton induced X-ray emission method for the elements P, K, Ca, Fe, Zn in the case of patients undergoing postoperative gamma irradiation following mastectomy. None of the concentration ratios were found to be influenced under the present conditions of irradiation.

Intraventricular administration of vasopressin and oxytocin effects the steady-state levels of serotonin, dopamine and norepinephrine in rat brain.

The effect of lysine-8-vasopressin (LVP) and oxytocin (OXT) has been studied on the steady-state level of serotonin (5-HT),dopamine(DA) and norepinephrine (NE) in various brain areas four hours after intraventricular microinjection of the neuropeptides. LVP (50 microU) diminished in content of 5-HT and NE in the mesencephalon, the content of 5-HT in the septum as well as the content of DA and NE in the dorsal hippocampus. OXT (500 microU), on the other hand, decreased the content of 5-HT, DA and Ne in the hypothalamus, as well as the content of DA in the mesencephalon. In contrast, the septal NW level was increased following OXT treatment. The data indicate that both LVP and OXT are capable of influencing the 5-HT and catecholamine levels as late as 4 h after their administration. Therefore, our findings support the previous notion that posterior pituitary neuropeptides affect behavioral processes via interacting with the cerebral monoaminergic neurotransmission.

Hereditary diabetes insipidus in rats. Altered cerebral indolamine and catecholamine metabolism.

Compared to heterozygous Brattleboro animals, homozygous (diabetes insipidus) rats exhibited higher steady-state levels of serotonin in the mesencephalon, septum and striatum. These differences disappeared upon the administration of pargyline, suggesting accumulation of serotonin. The norepinephrine level was higher in the mesencephalon, while the disappearance rate (alpha-met hyl-p-tyrosine) was accelerated in the septum and decreased in the hypothalamus. The lower striatal dopamine level was associated with a decreased disappearance rate. The data suggest that the altered monoamine metabolism might be associated with the known endocrine and behavioral disturbances of the homozygous rats.

A comparative study of DNA-induced transformants and spontaneous revertants of inositolless Neurospora crassa.

Inositolless (inl-) Neurospora crassa strains were treated with DNA (allo-DNA) of wild type N. Crassa. Hyphal fragments of a mycelial suspension of the N. Crassa ragged inl- strain used as recipient in our transformation experiments were found to consist of units containing 100--1000 nuclei. In this strain the inositol-independent (inl+) nuclei appearing after DNA treatment or by spontaneous reversion are present in the cytoplasm together with a large number of inl- nuclei. Thus, both transformation and reversion initially must result in heterokaryosis. Under appropriate conditions the inl- nuclei can be detected in the transformed and spontaneous inl+ phenotype revertant strains. Spontaneous revertants are usually characterized by the loss of their inl+ nuclei after transfers on inositol-supplemented medium. On minimal medium, the growth rate of transformed strains is significantly lower than that of spontaneous revertants. The inl+ gene appearing after DNA treatment or by spontaneous reversion is inherited as a trait bound to chromosomes. In crosses with the transformed strains, there is a significant increase in the number of non-Mendelian (6:2 and 5:3) tetrads in the inl locus.

Effect of ACTH and its fragment on brain catecholamines in intact and adrenalectomized rats.

Brain catecholamine metabolism was monitored by distribution of labelled noradrenaline (3H-NA) after intraventricular injection to intact and adrenalectomized rats. The adrenalectomy produced an increased disappearance rate of the labelled pool in the hypothalamus, hippocampus and neocortex. These changes could be prevented by hydrocortisone pretreatment. Painful stimuli resulted in an increased disappearance of the labelled pool in both intact and adrenalectomized rats. The implantation of hydrocortisone into the tuberoinfundibular region prevented the stress-induced changes of the catecholamine metabolism. Intraventricular administration of ACTH1-24 and ACTH4-10 produced a significant increase of the disappearance rate in different brain regions of adrenalectomized rats. The blocking of catecholamine synthesis by intraventricular injection of alpha-methyl-m-tyrosine resulted in a marked decrease of the labelled pool but did not prevent the ACTH-induced decrease of the tracer pool. On the other hand, the blocking of monoamine-oxydase activity by Pargyline led to a marked increase of the labelled pool but intraventricular administration of ACTH led to an increase of the disappearance rate. The mechanism of ACTH action on brain catecholamine metabolism is still obscure, however, an increased release of the NA to ACTH peptides is very likely in the light of the present observations.