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Extracellular vesicles (EVs) are small spherical particles surrounded by a membrane with an unusual lipid composition and a striking cholesterol/phospholipidic ratio. About 2000 lipid and 3500 protein species were identified in EVs secreted by different cell sources. EVs mediate cell to cell communication in proximity to or distant from the cell of origin. In particular, it was suggested that they represent modulators of multiple processes during pregnancy. The aim of this study was to identify the presence of EVs in canine amnion-derived cells (ASCs) culture and the expression of CD 59 on their surface. Amniotic membrane was collected in PBS with antibiotics added from 2 bitches during elective caesarean section. Cells culture was prepared and EVs were isolated. EVs were used to evaluate CD59 expression by flow cytofluorimetry. We found that the majority of EVs expressed CD59. Our results could increase the knowledge about the complex mechanisms that regulate the pregnancy in the bitch., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Background: Previous studies have indicated that correction of an established kyphosis in spine tuberculosis is both difficult and hazardous. There has not been any publication about evaluation of surgical correction of spine tuberculosis in Indonesia, despite of high incidence of spine tuberculosis cases. Therefore, we evaluated the outcome of kyphotic angle correction and neurological status after surgery of spinal tuberculosis patients for better understanding., Methods: Retrospectively, 96 patients with spinal tuberculosis that underwent operation in Soeharso Orthopedic Hospital from June 2016 to July 2019 were selected. Operation procedure includes laminectomy, debridement and posterior stabilization. We obtained plain x-ray of spine to evaluate the kyphotic deformity before and after surgery. We also examined neurological status of the patient before and after surgery., Results: The average pretreatment kyphotic angle in thoracic tuberculosis was 33.69° (range 8°-86°), which improved into a significant change to 13.27° (range 0°-56°). Correction angle was <25° in 34 people, 25°-50° in 17 people, and >50° in 4 people. While in the lumbar tuberculosis, it was 25.52° (range 6°-80°), and improved into 11.51° (range 2°-48°). Correction angle was <25° in 35 people, 25°-50° in 4 people, and >50° in 2 people. Improved neurological deficit was shown in 12% of patient with lumbar tuberculosis, and the rest had constant neurological deficit. While in thoracic tuberculosis found that 7% have improved neurological deficit and the rest is constant. None of them have worsen neurological status after the surgery., Conclusions: Surgical treatment for kyphotic deformity in patient with thoracolumbar tuberculosis are effective and safe, even in high corrective angle (>50°)., Competing Interests: Declaration of competing interest All authors have none to declare., (Copyright © 2021 Tuberculosis Association of India. Published by Elsevier B.V. All rights reserved.)

Objective: Cardiovascular (CV) morbidity is a well-established problem in systemic lupus erythematosus (SLE). Antimalarial (AM) therapy has been seen as a potential atheroprotective agent. The aim was to assess the impact of AM therapy on traditional and novel atherosclerosis (AT) biomarkers in patients with SLE., Methods: A search of MEDLINE, EMbase, and Cochrane library for studies evaluating the impact of AM on AT biomarkers in SLE was conducted. Data extraction included serum, functional and structural traditional and novel biomarkers. A narrative synthesis of the findings and a meta-analysis with random effects was conducted estimating mean differences (MD), OR, HR and 95% CIs., Results: The search strategy produced 148 articles, of which 64 were extracted for analysis. The MD in VLDL-cholesterol (-10.29, 95% CI -15.35, 5.24), triglycerides (-15.68, 95% CI -27.51, -3.86), and diastolic BP (-3.42, 95% CI -5.62, -1.23) differed significantly in patients on AM therapy compared with those without AM therapy. Patients on AM had a lower prevalence and incidence of diabetes mellitus than patients not on AM (HR: 0.39, 95% CI 0.17, 0.88). HCQ use was associated with lower blood pressure (BP) variability. Structural markers like carotid intima-media thickness (IMT), carotid plaque (CP) and coronary artery calcification (CAC) were not influenced by AM. For functional markers like endothelial and arterial stiffness the benefit was unclear. The GRADE approach showed a very low-to-low quality of evidence (QoE) per outcome., Conclusions: There is some evidence on the associations between AM therapy and some AT markers. However, the data on which this conclusion was based was of low to very low evidence., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Drosophila suzukii is an invasive pest that prefers to lay eggs in ripening fruits, whereas most closely related Drosophila species exclusively use rotten fruit as oviposition site. This behaviour is allowed by an enlarged and serrated ovipositor that can pierce intact fruit skin, and by multiple contact sensory systems (mechanosensation and taste) that detect the optimal egg-laying substrates. Here, we tested the hypothesis that bristles present in the D. suzukii ovipositor tip contribute to these sensory modalities. Analysis of the bristle ultrastructure revealed that four different types of cuticular elements (conical pegs type 1 and 2, chaetic and trichoid sensilla) are present on the tip of each ovipositor plate. All of them have a poreless shaft and are innervated at their base by a single neuron that ends in a distal tubular body, thus resembling mechanosensitive structures. Fluorescent labelling in D. suzukii and D. melanogaster revealed that pegs located on the ventral side of the ovipositor tip are innervated by a single neuron in both species. RNA-sequencing profiled gene expression, notably sensory receptor genes of the terminalia of D. suzukii and of three other Drosophila species with changes in their ovipositor structure (from serrated to blunt ovipositor: Drosophila subpulchrella, Drosophila biarmipes and D. melanogaster). Our results revealed few species-specific transcripts and an overlapping expression of candidate mechanosensitive genes as well as the presence of some chemoreceptor transcripts. These experimental evidences suggest a mechanosensitive function for the D. suzukii ovipositor, which might be crucial across Drosophila species independently from ovipositor shape., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Background and Aims: Non-invasive surrogates of cardiovascular (CV) disease such as endothelial dysfunction (ED) and peripheral arterial stiffness (AS) have been evaluated in systemic lupus erythematosus (SLE) patients. The aim of this study was to systematically review and meta-analyze reports of cardiovascular disease (CVD) in SLE patients, as measured by ED and AS., Methods: Studies analyzing the relationship of SLE with ED (flow-mediated dilatation [FMD], nitroglycerin-mediated dilatation [NMD] and peripheral arterial tonometry [PAT]) and AS (augmentation index [AIx], pulse wave velocity [PWV]) were systematically searched for in PubMed, Cochrane library, EMBASE, VHL, SciELO and Web of Science databases. Inclusion criteria included peer-review and English language. Mean differences (MD) and 95% confidence intervals (CIs) were estimated using the random effect model. The study was registered with PROSPERO, number CRD42019121068., Results: The meta-analysis included 49 studies. FMD data from 18 studies including 943 SLE subjects (mean age = 38.71 [95%CI 36.21, 41.21] years) and 644 unaffected controls (mean age = 38.63 [95%CI 36.11, 41.15] years) were included. When compared with unaffected controls, FMD in SLE subjects was decreased by 4.3% (95%CI: -6.13%, -2.47%): p < 0.001). However, NMD did not significantly differ between SLE patients and controls (MD = - 2.68%; 95% CI -6.00, 0.62; p = 0.11). A significantly increased AS between SLE patients and controls according to overall PWV (MD = 1.12 m/s; 95% CI 0.72-1.52; p < 0.001) was observed, but not for the brachial-ankle PWV. AIx was also increased in SLE patients compared with healthy controls (MD = 4.55%; 95% CI 1.48-7.63; p = 0.003)., Conclusions: Overall, SLE patients showed impaired FMD, an independent predictor of CV events. There was a higher degree of AS in SLE patients compared with controls. ED and AS in SLE should be considered when planning preventive strategies and therapies., Competing Interests: Declaration of competing interest The authors declared they do not have anything to disclose regarding conflict of interest with respect to this manuscript., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Patients with indeterminate thyroid nodules (Bethesda III and IV) are often treated with diagnostic lobectomy, which in most cases represents an overtreatment. A reliable rule-out molecular test could spare patients unnecessary surgery. Stained smears of 88 indeterminate thyroid nodules with histologic diagnosis of follicular-patterned tumors were selected: 34 follicular adenomas (FAs), 34 follicular variant papillary thyroid carcinomas (FVPTCs), and 20 noninvasive follicular neoplasms with papillary-like nuclear features (NIFTPs). The expression level of 126 genes was measured by digital counting. Mutation testing was performed for the main gene mutations and fusions. Performance of gene expression and mutation tests was calculated by receiver operating characteristic analysis. The gene expression model showed an area under the curve (AUC) of 88%, with 91% negative predictive value in FAs and FVPTCs only. Part of NIFTPs was labeled as benign, and part was labeled as malignant; thus, the classifier performance worsened. Two FAs (5.9%), eight NIFTPs (40%), and 22 FVPTCs (64.7%) were mutation positive. Mutation testing AUC was 79% in FAs and FVPTCs, and decreased by including NIFTPs. This gene expression-based test was feasible in thyroid-stained smears, showed higher AUC than mutation test, and had a high negative predictive value-making it a good candidate as a rule-out test for indeterminate thyroid cytology. NIFTPs have a heterogeneous phenotype, and their preoperative diagnosis requires further investigation., (Copyright © 2020 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.)

Objective: Obesity has a great impact on cardiovascular morbidity and mortality, the treatment of this pathological state is important given the significant health consequences. Lifestyle and behaviour changes play a significant role in weight management. The purpose of this study was to investigate the impact of an intensive multidisciplinary lifestyle intervention on well-known atherogenic factors in a group of overweight and obese subjects., Methods: A total of 44 people with overweight/obesity underwent a lifestyle intervention based on nutritional education, psychological support and a 3-month exercise training program with a frequency of twice a week. Several anthropometric and biochemical parameters were measured before and after the lifestyle intervention., Results: Lifestyle intervention led to a significant reduction in metabolic profile including body mass index (BMI), waist circumference, systolic and diastolic blood pressure, plasma glucose, and plasma triglycerides. These reductions were also accompanied by a significant increase in maximal oxygen consumption and muscle strength. Furthermore, paraoxonase and lactonase activities and the concentration of Apoliproteins A1 (APO A1) were significantly increased and the serum levels of oxLDL reduced without any changes in the circulating levels of LDL and HDL., Conclusion: In conclusion, our study suggests that an intensive lifestyle intervention in obese subjects promotes a series of beneficial antiatherogenic changes which included increased enzyme activites of paraoxonase and lactonase, concentration of Apoliproteins A1 and decreased circulating levels of oxLDL., (Copyright © 2016 Asia Oceania Association for the Study of Obesity. Published by Elsevier Ltd. All rights reserved.)

The idea of possible involvement of the aryl hydrocarbon receptor (AhR) in transplant tolerance can be traced back >30 years, when very low doses of dioxin-the most potent AhR ligand-were found to markedly reduce the generation of cytotoxic T lymphocytes in response to alloantigen challenge in vivo. AhR is a ligand-activated transcription factor that is activated by dioxins and other environmental pollutants. We now know that AhR can bind a broad variety of activating ligands that are disparate in nature, including endogenous molecules and those formed in the gut from food and bacterial products. Consequently, in addition to its classical role as a toxicological signal mediator, AhR is emerging as a transcription factor involved in the regulation of both innate and adaptive immune responses in various immune cell types, including lymphocytes and antigen-presenting cells (APCs). Allograft rejection is mostly a T cell-mediated alloimmune response initiated by the recognition of alloantigens presented by donor and recipient APCs to recipient CD4(+) and CD8(+) T cells. Based on those findings, AhR may function as a critical sensor of outside and inside environments, leading to changes in the immune system that may have relevance in transplantation., (© Copyright 2016 The American Society of Transplantation and the American Society of Transplant Surgeons.)

Direct thermal desorption coupled with the Gas Chromatography-Mass Spectrometry (TDU-CIS4-GC-MS) technique applied to powdered milk was used as a novel approach for the characterization and quantification, as relative abundance, of volatile organic compounds (VOCs). The aim of this study was to exploit the potential applications and setup conditions of the CIS4-TDU-GC-MS technique for the identification of oxidized or non-oxidized samples of powdered milk for infant nutrition, subjected to accelerated aging through the changes in their VOCs profile over time. Thermal desorption at 30°C and subsequent cryotrapping at -40°C with a Gerstel Liner-in-Liner system allowed a direct thermal desorption and cryotrapping of VOCs without any memory effect, thus avoiding sample preparation and contamination. The analyses led to the identification of several characteristic off-flavors in the oxidized samples, which were used as molecular markers to discriminate samples with different oxidation degrees. Finally, VOC contaminants possibly present in the packaging can also be identified with this technique., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Dendritic cells (DCs) are specialized antigen-presenting cells with a bipolar nature. Depending on environmental factors, DCs will promote either inflammatory or anti-inflammatory effects. Lipopolysaccharide (LPS), a ligand of Toll-like receptor (TLR)4 and a most potent proinflammatory stimulus, is responsible for complex signaling events in different cell types, including DCs. LPS effects range from protective inflammation-capable of counteracting growth and dissemination of gram-negative bacteria - to hyperacute detrimental responses, as it occurs in endotoxic shock. Consistent with the plasticity of TLR4 signaling, a low dosage of LPS will induce a regulatory response capable of protecting mice against a subsequent, otherwise lethal challenge ('endotoxin tolerance'). By examining CD11c(+) DCs ('conventional' DCs, or cDCs), we investigated whether DC flexibility in promoting either inflammation or tolerance can be differentially affected by single vs. repeated exposure to LPS in vitro. cDCs stimulated twice with LPS expressed high levels of indoleamine 2,3-dioxygenase 1 (IDO1) - one of the most effective mediator of anti-inflammatory activity by DCs - and of TGF-β, an immunoregulatory cytokine capable of upregulating IDO1 expression and function. In contrast, a single exposure to LPS failed to upregulate IDO1, and it was instead associated with high-level production of IL-6, a cytokine that promotes inflammation and proteolysis of IDO1. When adoptively transferred in vivo, only cDCs on double endotoxin exposure greatly improved the outcome of an otherwise lethal LPS challenge. The protective effect required that the transferred cDCs be fully competent for IDO1 and the host for TGF-β production. Thus cDCs, conditioned by LPS in vitro to mimic an endotoxin-tolerant state, can protect recipients from endotoxic shock, pointing to adoptive transfer of tolerance as a new option for controlling potentially harmful responses to TLR4 signaling., (Copyright © 2014 Elsevier GmbH. All rights reserved.)

Objective: We reviewed the surgical complications from our recent experience in vascular and tumor patients who underwent anterior clinoidectomy through the lateral supraorbital (LSO) approach., Methods: Between June 2007 and January 2011, a total of 82 patients with neoplastic and vascular lesions underwent anterior clinoidectomy by the senior author (J.H.) through the LSO approach. We analyzed the operative videos paying particular attention to the surgical technique used for removal of the anterior clinoid process (ACP) and compared the microsurgical nuances to postoperative complications related to anterior clinoidectomy., Results: Forty-five patients were treated for aneurysms; 35 patients for intraorbital, parasellar, and suprasellar tumors; and 2 patients for carotid-cavernous fistulas. Intradural anterior clinoidectomy was performed in 67 (82%) cases; in 15 (18%) cases an extradural approach was used. In 51 (62%) cases, ACP was removed completely, whereas in the remaining 31 (38%) a tailored anterior clinoidectomy was performed. Four (5%) patients had new postoperative visual deficits and 3 (4%) experienced a worsening of preoperative visual deficits. Twelve (15%) patients improved their preoperative visual deficits after intradural anterior clinoidectomy. Ultrasonic bone device is a useful tool but may damage the optic nerve when performing anterior clinoidectomy. There was no mortality in our series., Conclusion: Anterior clinoidectomy can be performed through an LSO approach with a safety profile that is comparable to other approaches. Ultrasonic bone dissector is a useful tool but may lead to injury of the optic nerve and should be used very carefully in its vicinity., (Copyright © 2012. Published by Elsevier Inc.)

Objective: To demonstrate that anterior clinoidectomy is possible through the lateral supraorbital (LSO) approach, and that extent of the clinoidectomy is tailored according to the lesion. We reviewed our recent experience on patients with vascular and tumor who underwent anterior clinoidectomy through the LSO approach., Methods: Between June 2007 and January 2011, 82 patients with neoplastic and vascular lesions underwent anterior clinoidectomy by the senior author (J.H.) with the LSO approach. We retrospectively analyzed the surgical videos and the microsurgical techniques of anterior clinoidectomy., Results: Forty-five patients were treated for aneurysms, 35 patients for intraorbital, parasellar and suprasellar tumors, and 2 patients presented with carotid-cavernous fistula. Intradural anterior clinoidectomy was performed in 67 patients (82%); in 15 patients (18%) extradural anterior clinoidectomy was used. A minimal removal of the anterior clinoid process (ACP) was performed in 5 patients, in 8 patients a partial clinoidectomy was performed, in 18 patients a subtotal removal of the ACP was needed, and in 51 patients, the entire ACP was removed. There was no operation-related mortality in the series., Conclusions: A tailored anterior clinoidectomy is useful and can be performed through the LSO approach. Intradural visualization of the internal carotid artery and optic nerve is mandatory for the exact anatomic orientation and safe anterior clinoidectomy. We recommend intradural anterior clinoidectomy for all vascular and most neoplastic lesions., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Objective: To report the case of a young patient with juvenile nasopharyngeal angiofibroma (JNA) (Fisch grade IVb) operated on in a semisitting position., Methods: A 17-year-old boy complained of nasal obstruction and pain in the left periorbital area in April 2007. Magnetic resonance imaging (MRI) showed grade IVb JNA with invasion of the anterior cranial fossa, the cavernous sinuses, and the chiasm. Resections of the tumor through bifrontal craniotomy in June 2007 and through lateral rhinotomy in January 2008, both performed in the supine position, were complicated by excessive bleeding., Results: In February 2008, the lesion was removed through a transfacial approach in a semisitting position after transnasal and percutaneous embolization. The vascular support to the tumor was reduced by using high-power bipolar forceps, monopolar diathermy, and intratumoral injection of fibrin glue. The main part of the lesion was successfully removed., Conclusions: The combination of preoperative embolization (transnasal, percutaneous, endovascular), semisitting positioning of the patient, intraoperative injection of fibrin glue, and high-power bipolar forceps and monopolar diathermy is helpful in reducing surgical bleeding and enabling safe resection of JNA., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Objective: To present a summary of anesthetic considerations for use of the sitting position in procedures to remove lesions of the occipital and suboccipital regions, with a special reference to the Helsinki experience with more than 300 operations in 1997-2007, and a retrospective study evaluating the incidence of venous air embolism (VAE) and hemodynamic stability in patients operated in the steep sitting position., Methods: Anesthesiology reports of 72 patients with a mean (± standard deviation [SD]) age of 33 years ± 18 treated by the senior author (J.H.) for pineal region tumors using the infratentorial supracerebellar approach in the sitting position during an 11-year period were retrospectively reviewed for the incidence of VAE and hemodynamic stability., Results: In the sitting position, median systolic blood pressure changed -8 (-95 to +50) mm Hg without alteration in heart rate. Based on patient records, the incidence of VAE was 19% (14 of 72 patients). In five patients, end-tidal carbon dioxide (ETCO(2)) decreased more than 0.7 kPa (5.25 mm Hg), possibly indicating VAE. Comparing patients with and without VAE, no differences in change of blood pressure, heart rate, or amount of administered vasoactive agents were observed. Postoperative duration of ventilator treatment and hospital stay were similar in patients with and without VAE. No signs of arterial embolization were seen postoperatively., Conclusions: The sitting position is associated with risk for hypotension. The same surgical approach and procedure does not exclude the occurrence of VAE. In this study, the unaltered hemodynamics in patients during VAE indicates relatively small VAE. Possible explanations for this are early recognition of air leakage and good cooperation between the surgical and anesthesia teams., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Objective: Anterior choroid artery aneurysms (AChAAs) constitute 2%-5% of all intracranial aneurysms. They are usually small, thin walled with one or several arteries originating at their base, and often associated with multiple aneurysms. In this article, we review the practical microsurgical anatomy, the preoperative imaging, surgical planning, and the microneurosurgical steps in the dissection and the clipping of AChAAs., Methods: This review, and the whole series on intracranial aneurysms (IAs), are mainly based on the personal microneurosurgical experience of the senior author (J.H.) in two Finnish centers (Helsinki and Kuopio) that serve, without patient selection, the catchment area in Southern and Eastern Finland., Results: These two centers have treated more than 10,000 patients with IAs since 1951. In the Kuopio Cerebral Aneurysm Database of 3005 patients with 4253 IAs, 831 patients (28%) had altogether 980 internal carotid artery (ICA) aneurysms, of whom 95 patients had 99 (2%) AChAAs. Ruptured AChAAs, found in 39 patients (41%), with median size of 6 mm (range = 2-19 mm), were associated with intracerebral hematoma (ICH) in only 1 (3%) patient. Multiple aneurysms were seen in 58 (61%) patients., Conclusions: The main difficulty in microneurosurgical management of AChAAs is to preserve flow in the anterior choroid artery originating at the base and often attached to the aneurysm dome. This necessitates perfect surgical strategy based on preoperative knowledge of 3 dimensional angioarchitecture and proper orientation during the microsurgical dissection., (Copyright © 2010 Elsevier Inc. All rights reserved.)

The role of Na(+)-Ca(2+) exchanger (NCX) in vascular endothelium is still matter of debate. Depending on both the endothelial cell (EC) type and the extracellular ligand, NCX has been shown to operate in either the forward (Ca(2+) out)- or the reverse (Ca(2+) in)-mode. In particular, acetylcholine (Ach) has been shown to promote Ca(2+) inflow in the intact endothelium of excised rat aorta. Herein, we assessed the involvement of NCX into the Ca(2+) signals elicited by ATP in such preparation. Removal of extracellular Na(+) (0Na(+)) causes the NCX to switch into the reverse-mode and induced an increase in intracellular Ca(2+) concentration ([Ca(2+)](i)), which disappeared in the absence of extracellular Ca(2+), and in the presence of benzamil, which blocks both modes of NCX, and KB-R 7943, a selective inhibitor of the reverse-mode. ATP induced a transient Ca(2+) signal, whose decay was significantly prolonged by 0Na(+), benzamil, DCB, and monensin while it was unaffected by KB-R 7943. Notably, lowering extracellular Na(+) concentration increased the sensibility to lower doses of ATP. These date suggest that, unlike Ach-stimulated ECs, NCX promotes Ca(2+) extrusion when the stimulus is provided by ATP in intact endothelium of rat aorta. These data show that, within the same preparation, NCX operates in both modes, depending on the chemical nature of the extracellular stimulus., (2010 Elsevier Inc. All rights reserved.)

Background: Because of the complex topographic anatomical relationship between vascular, dural and bone structures, paraclinoid aneurysms, especially those of larger size, remain a great challenge for vascular neurosurgeons. We present our microneurosurgical experience of 51 consecutive patients with large and giant paraclinoid aneurysms to scrutinize our personal strategies related to surgical treatment., Methods: Fifty-one patients with large or giant paraclinoid underwent micorneurosurgical aneurysm treatment. Operative strategies were planned according to preoperative state-of-the-art imaging studies, and a pterional-transsylvian approach was routinely used. Proximal control of the internal carotid artery (ICA) was achieved by exposure of the cervical portion of the vessel. Intraoperative electroencephalogram and somatosensory evoked potential monitoring, indocyanine green (ICG) videoangiography and/or microvascular Doppler ultrasonography (MDU) were regularly used. A postoperative digital subtraction angiography or computed tomography angiography was performed to verify the efficacy of treatment., Results: Forty-three large and giant paraclinoid aneurysm necks (84%) were directly clipped, seven unclippable aneurysms (14%) were trapped with extra-intracranial high-flow revascularization, and one aneurysm (2%) was treated with only ICA proximal Hunterian ligation. Two patients (4%) died in the early postoperative period. In 84% of the patients, the Glasgow Outcome Scale score was 4 or 5 at discharge. At the 6-month follow-up examination, the Rankin Outcome Scale score was 0-2 in 90% of patients., Conclusions: Temporary parent vessel occlusion, retrograde suction decompression, endoaneurysmectomy, parent vessel clip reconstruction, and bypass vascular anastomosis are essential techniques to treat complex paraclinoid aneurysms. The combined use of electrophysiological monitoring, MDU, intraoperative ICG videoangiography, and endoscopy can substantially improve microsurgical outcome., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Paraoxonases (PONs) are a small family of antioxidant enzymes whose antiatherogenic activity is well known. The aim of the present study was the evaluation of the effects of moderate aerobic training on their expression using a rat model. In order to discriminate between PON1 and PON3 enzymatic activity, we took advantage of some differences in their substrate preferences. PON1 and PON3 enzymatic activities and their protein levels were analyzed in plasma and in liver microsomes, and their mRNA levels in the liver. Exercise training did not affect PON1 expression or enzymatic activity but increased PON3 mRNA, protein levels, and enzymatic activity. Training also induced variations in plasma membrane composition, including an increase in polyunsaturated and a decrease in mono- and di-unsaturated fatty acids. On the other hand, acute exercise inhibited PON activities while increasing PON3 protein content in liver microsomes and reversing the relative composition in mono-, di-, and poly-unsaturated fatty acids, suggesting that physical stress, by altering membrane composition, may impair PON release from liver membranes. In conclusion, we documented, for the first time, the presence of PON3 in rat serum and, notably, found that the upregulation of PON3, rather than PON1, appears to be associated with physical training.

The inner wall of excised rat aorta was scraped by a microelectrode and Ca2+ signals were investigated by fluorescence microscopy in endothelial cells (ECs) directly coupled with injured cells. The injury caused an immediate increase in the intracellular Ca2+ concentration ([Ca2+]i), followed by a long-lasting decay phase due to Ca2+ influx from extracellular space. The immediate response was mainly due to activation of purinergic receptors, as shown by the effect of P2X and P2Y receptors agonists and antagonists, such as suramin, alpha,beta-MeATP, MRS-2179 and 2-MeSAMP. Inhibition of store-operated Ca2+ influx did not affect either the peak response or the decay phase. Furthermore, the latter was: (i) insensitive to phospholipase C inhibition, (ii) sensitive to the gap junction blockers, palmitoleic acid, heptanol, octanol and oleamide, and (iii) sensitive to La3+ and Ni2+, but not to Gd3+. Finally, ethidium bromide or Lucifer Yellow did not enter ECs facing the scraped area. These results suggest that endothelium scraping: (i) causes a short-lasting stimulation of healthy ECs by extracellular nucleotides released from damaged cells and (ii) uncouples the hemichannels of the ECs facing the injury site; these hemichannels do not fully close and allow a long-lasting Ca2+ entry.

In several species of hymenopteran parasitoids of the superfamilies of Ichneumonoidea and Platygastroidea, the membrane enveloping the parasitoid embryo dissociates at hatching into a number of cells, called teratocytes, which autonomously develop in the host haemolymph. In this work we report for Encarsia berlesei and Encarsia citrina (Hymenoptera: Chalcidoidea), the dissociation of the extraembryonic membrane into cells whose morphological and embryological features correspond to those of teratocytes. In E. berlesei the membrane dissociated at hatching into 4-9 larger cells (100 microm diameter) and about 10 smaller cells (60 microm), which scarcely doubled their size during maturation. In E. citrina the membrane dissociated into five large cells (250 microm) which did not grow appreciably. Ultrastructural investigation of the dissociated cells in E. berlesei revealed that their surface was covered by microvilli, whose density and length increased from the egg stage to the 12 h following hatching. During the same period, rough endoplasmic reticulum evolved from a parallel profile to that of the cisternal type, while abundant vesicles represented the dominant cytological feature. The ploidy level of these cells ranged between 8c and 140c at hatching, but increased to 40c-350c at maturation. These findings provide the first clear evidences for the presence of teratocytes in the superfamily Chalcidoidea.

The present study looks at possible changes in the activity of acetylcholinesterase (AChE) in tissues (brain and white muscle) of the Mediterranean bony fish Sparus auratus after a 20 days exposure to sublethal concentrations (0.1 or 0.5 ppm) of copper in the marine water and on control untreated animals. The trials also included measurements of Cu concentration in the tissues to evaluate possible metal accumulation. Moreover, sedimentation analysis as well as V(max) and K(m) determination were carried out in tissue extracts of Cu-exposed or control animals. V(max) and K(m) were also determined with or without addition of Cu(2+) in the assay. No Cu accumulation occurred in brain and muscle after Cu exposure. AChE showed in both tissues a molecular polymorphism with putative globular (G) and asymmetric (A) forms. Cu exposition led to an increased specific activity and improved catalytic efficiency of AChE in brain and muscle, seemingly regarding G forms. The increase in catalytic efficiency also resulted from the in vitro assay with tissue extracts and Cu(2+) addition. The higher AChE activity and catalytic efficiency in both tissues after Cu exposition and without metal accumulation, suggests an increase of free Cu aliquot into the cells, likely due to mechanisms of metal homeostasis.

The present study regards possible changes in the activity of glyoxalase system enzymes (glyoxalase I, GI, and glyoxalase II, GII) in tissues (brain, liver and white muscle) of the mediterranean bony fish Sparus auratus after a 20 days exposure to sublethal concentrations (0.1 or 0.5 ppm) of Cu in the marine water and on control untreated animals. The experiments also included measurements of copper concentration in the tissues, as well as of lactate dehydrogenase (LDH) activity, to evaluate possible Cu accumulation and changes in glycolytic activity respectively. Cu accumulation only occurs in the liver. GI, GII and LDH activities kept unchanged in the brain after copper exposure. GI activity in liver and muscle of copper-exposed animals decreases probably for a slackening in the glycolytic rate, as suggested by the lowering of LDH activity. GII activity remains unchanged or increases (liver extract, 0.5 ppm of Cu), maybe to safeguard enough cellular levels of GSH., (Copyright 2002 Elsevier Science B.V.)

Three forms of acetylcholinesterase (AChE) were detected in samples of the bivalve mollusc Mytilus galloprovincialis collected in sites of the Adriatic sea. Apart from the origin of the mussels, two spontaneously soluble (SS) AChE occur in the hemolymph and represent about 80% of total activity, perhaps hydrolyzing metabolism-borne choline esters. These hydrophilic enzymes (forms A and B) copurified by affinity chromatography (procainamide-Sepharose gel) and were separated by sucrose gradient centrifugation. They are, respectively, a globular tetramer (11.0-12.0 S) and a dimer (6.0-7.0 S) of catalytic subunits. The third form, also purified from tissue extracts by the same affinity matrix, proved to be an amphiphilic globular dimer (7.0 S) with a phosphatidylinositol tail giving cell membrane insertion, detergent (Triton X-100, Brij 96) interaction and self-aggregation. Such an AChE is likely functional in cholinergic synapses. All three AChE forms show a good substrate specificity and are inactive on butyrylthiocholine. Studies with inhibitors showed low inhibition by eserine and paraoxon, especially on SS forms, high sensitivity to 1,5-bis(4-allyldimethylammoniumphenyl)-pentan-3-one dibromide (BW284c51) and no inhibition with propoxur and diisopropylfluorophosphate (DFP). The ChE forms in M. galloprovincialis are possibly encoded by different genes. Some kinetic features of these enzymes suggest a genetic polymorphism.

5-fluorouracil (5-FU) is still the most effective cytotoxic agent for the treatment of human colorectal cancer. Response rates, however, vary between 5-20%. One attempt to improve the effect of 5-FU is through biomodulation. We have previously found the somatostatin analogue, SMS 201.995 (Sandostatin, Sandoz), to inhibit both the in-vitro and in-vivo growth of some human colon cancer cell lines. It may act specifically by means of receptors on the surface of tumour cells, or by reducing the concentration of some growth factors. We report that, when 5-FU at 0.125 and 0.25 micrograms/ml was combined with SMS 201.995 at 10(-12) x 2 to 10(-8) x 2M, an enhanced inhibition of in-vitro growth of two human colorectal cancer cell lines (C170 and LIM 1215) was achieved. Effects were measured using [3H]-thymidine uptake and by a colorimetric assay of cellular respiration (MTT, Promega, Sydney). SMS 201.995 alone has minimal inhibitory effects, whilst 5-FU alone shows inhibition as low as 39.6% of control. When 5-FU was then combined with SMS 201.995, a 10-30% inhibition occurred compared to the 5-FU control. The combination of 5-FU and SMS 201.995 may be a useful method of improving response to human colorectal cancer therapy.