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This study evaluated the relationship between body size and digestive tract characteristics of the important predatory freshwater fish Hoplias malabaricus, which is widely distributed in South America. The allometric coefficients were calculated for the mass and standard length ( LS) relationships for two different LS groups: (1) between 20 and 100 mm (characterized as insectivores) and (2) >100 mm (characterized as piscivores). Differential growth measured from the allometric coefficient, b, between the insectivore ( b < 3) and the piscivore ( b > 3) groups was detected. Anterior intestine length and pyloric caeca zone length showed significant differences between groups. Two complementary hypotheses were developed to explain the differential growth: (1) H. malabaricus has a digestive tract adapted to a piscivorous diet, which is independent of its ontogenetic stage of development, and (2) the negative allometry observed in group 1 individuals agrees with a general behavioural strategy, allowing individuals to grow in LS during a shorter period of time. [ABSTRACT FROM AUTHOR]

Plasma malondialdehyde-like material (MDA-LM) was evaluated in 138 normal subjects and in a group of 57 stroke patients using a modification of the method of Smith et al. (1976). The basal level of MDA-LM in the control group was 35 mumol/l with a range of 22-50 mumol/l. Values above 50 mumol/l were found in 80% of the patients suffering from subarachnoid haemorrhage, in 68% of those with cerebral thrombosis, and in 17% with transient ischaemic attacks. None of the patients with cerebral embolism, intracerebral haematoma, or lacunar infarct had values above 50 mumol/l. Significant statistical differences were found between the control group and all the patients except those with lacunar infarcts. [ABSTRACT FROM PUBLISHER]

Nonfluorinated polymers have been widely proposed to replace Nafion as raw materials for redox flow battery ion-exchange membranes. Hereby, block copolymers based on polysulfone (PSU) and polyphenylsulfone (PPSU) are synthesized and employed as precursors of membranes for vanadium redox flow batteries. A series of copolymers with varying molar proportions of PSU (75/25, 60/40, 50/50 mol %) were prepared. The 60/40 and 75/25 copolymers exhibit concentrated sulfonic groups predominantly in the PSU unit, favoring the formation of hydrophobic and hydrophilic domains. The 50/50 copolymer presents a balanced degree of sulfonation between the two units, leading to a homogeneous distribution of sulfonic groups. An ex situ study of these materials comprising vanadium ion permeability and chemical and mechanical stability was performed. The best performance is achieved with 50/50 membranes, which exhibited performance comparable to commercial Nafion membranes. These results signify a promising breakthrough in the pursuit of high-performance, sustainable membranes for next-generation VRFBs., Competing Interests: The authors declare no competing financial interest., (© 2024 The Authors. Published by American Chemical Society.)

Chitosan (CS)-based scaffolds loaded with Pinus radiata extract bark (PE) and grape seed extract (GSE) were successfully developed for wound dressing applications. The effects of incorporating GSE and PE in CS scaffolds were investigated in relation to their physicochemical and biological properties. All scaffolds exhibited porous structures with the ability to absorb more than 70 times their weight when contacted with blood and phosphate buffer solution. The incorporation of GSE and PE into the CS scaffolds increased their blood absorption ability and degradation rates over time. All scaffolds showed a clotting ability above 95%, with their surfaces being favorable for red blood cell attachment. Both GSE and PE were released from the CS scaffolds in a sustained manner. Scaffolds loaded with GSE and PE inhibited the bacterial activity of S. aureus and E. coli by 40% and 44% after 24 h testing. In vitro cell viability studies demonstrated that all scaffolds were nontoxic to HaCaT cells. Importantly, the addition of GSE and PE further increased cell viability compared to that of the CS scaffold. This study provides a new synthesis method to immobilize GSE and PE on CS scaffolds, enabling the formation of novel material platforms with a high potential for wound dressing applications.

The La Palma 2021 volcanic eruption was the first subaerial eruption in a 50-year period in the Canary Islands (Spain), emitting ~1.8 Tg of sulphur dioxide (SO 2 ) into the troposphere over nearly 3 months (19 September-13 December 2021), exceeding the total anthropogenic SO 2 emitted from the 27 European Union countries in 2019. We conducted a comprehensive evaluation of the impact of the 2021 volcanic eruption on air quality (SO 2 , PM 10 and PM 2.5 concentrations) utilising a multidisciplinary approach, combining ground and satellite-based measurements with height-resolved aerosol and meteorological information. High concentrations of SO 2 , PM 10 and PM 2.5 were observed in La Palma (hourly mean SO 2 up to ~2600 μg m -3 and also sporadically at ~140 km distance on the island of Tenerife (> 7700 μg m -3 ) in the free troposphere. PM 10 and PM 2.5 daily mean concentrations in La Palma peaked at ~380 and 60 μg m -3 . Volcanic aerosols and desert dust both impacted the lower troposphere in a similar height range (~ 0-6 km) during the eruption, providing a unique opportunity to study the combined effect of both natural phenomena. The impact of the 2021 volcanic eruption on SO 2 and PM concentrations was strongly influenced by the magnitude of the volcanic emissions, the injection height, the vertical stratification of the atmosphere and its seasonal dynamics. Mean daily SO 2 concentrations increased during the eruption, from 38 μg m -3 (Phase I) to 92 μg m -3 (Phase II), showing an opposite temporal trend to mean daily SO 2 emissions, which decreased from 34 kt (Phase I) to 7 kt (Phase II). The results of this study are relevant for emergency preparedness in all international areas at risk of volcanic eruptions; a multidisciplinary approach is key to understand the processes by which volcanic eruptions affect air quality and to mitigate and minimise impacts on the population., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Using in vitro and in vivo models, this study investigated the hemostatic potential to control bleeding of both unloaded gelatin-graphene oxide aerogels and the same loaded with proanthocyanidins (PAs) from Vitis vinifera grape skin extract. Our results showed that the physicochemical and mechanical properties of the aerogels were not affected by PA inclusion. In vitro studies showed that PA-loaded aerogels increased the surface charge, blood absorption capacity and cell viability compared to unloaded ones. These results are relevant for hemostasis, since a greater accumulation of blood cells on the aerogel surface favors aerogel-blood cell interactions. Although PAs alone were not able to promote hemostasis through extrinsic and intrinsic pathways, their incorporation into aerogels did not affect the in vitro hemostatic activity of these composites. In vivo studies demonstrated that both aerogels had significantly increased hemostatic performance compared to Spongostan TM and gauze sponge, and no noticeable effects of PA alone on the in vivo hemostatic performance of aerogels were observed; this may have been related to its poor diffusion from the aerogel matrix. Thus, PAs have a positive effect on hemostasis when incorporated into aerogels, although further studies should be conducted to elucidate the role of this extract in the different stages of hemostasis.

In this study, graphene oxide (GO)-based aerogels cross-linked with chitosan (CS), gelatin (GEL), and polyvinyl alcohol (PVA) were characterized and their hemostatic efficiencies both in vitro and in vivo were investigated and compared to commercial materials (ChitoGauze®XR and Spongostan™). All aerogels exhibited highly porous structures and a negative surface charge density favorable to their interaction with blood cells. The in vitro studies showed that all aerogels coagulated >60 % of the blood contained in their structures after 240 s of the whole-blood clotting assay, the GO-CS aerogel being the one with the highest blood clotting. All aerogels showed high hemocompatibility, with hemolytic rates <5 %, indicating their use as biomaterials. Among them, the GO-GEL aerogel exhibited the lowest hemolytic activity, due possibly to its high GEL content compared to the GO amount. According to their blood clotting activity, aerogels did not promote coagulation through extrinsic and intrinsic pathways. However, their surfaces are suitable for accelerating hemostasis by promoting alternative routes. All aerogels adhered platelets and gathered RBCs on their surfaces, and in addition the GO-CS aerogel surface also promoted the formation of filamentous fibrin networks adhered on its structure. Furthermore, in vivo evaluations revealed that all aerogels significantly shortened the hemostatic times and reduced the blood loss amounts compared both to the Spongostan™ and ChitoGauze®XR commercial materials and to the gauze sponge (control group). The hemostatic performance in vitro and in vivo of these aerogels suggests that they could be used as hemostats for controlling profuse bleedings., (Copyright © 2022 Elsevier B.V. All rights reserved.)

In this study, poly(ε-caprolactone) (PCL)/gelatin (GEL) electrospun nanofibers loaded with two different concentrations of Pinus radiata bark extracts (PEs) were fabricated via electrospinning for wound healing applications. The effects of incorporating PE into PCL/GEL electrospun nanofibers were investigated regarding their physicochemical properties and in vitro biocompatibility. All electrospun nanofibers showed smooth, uniform, and bead-free surfaces. Their functional groups were detected by ATR-FTIR spectroscopy, and their total phenol content was measured by a Folin-Ciocalteu assay. With PE addition, the electrospun nanofibers exhibited an increase in their wettability and degradation rates over time and a decrease in their tensile stress values from 20 ± 4 to 8 ± 2 MPa for PCL/GEL and PCL/GEL/0.36%PE samples, respectively. PE was also released from the fibrous mats in a rather controlled fashion. The PCL/GEL/0.18%PE and PCL/GEL/0.36%PE electrospun nanofibers inhibited bacterial activity at around 6 ± 0.1% and 23 ± 0.3% against E. coli and 14 ± 0.1% and 18 ± 0.2% against S. aureus after 24 h incubation, respectively. In vitro cell studies showed that PE-loaded electrospun nanofibers enhanced HaCaT cell growth, attachment, and proliferation, favoring cell migration towards the scratch area in the wound healing assay and allowing a complete wound closure after 72 h treatment. These findings suggested that PE-loaded electrospun nanofibers are promising materials for antibiotic-free dressings for wound healing applications.

Aerogels based on gelatin and graphene oxide (GO) were synthesized by microwave-assisted reactions, incorporating grape skin extracts -high in proanthocyanidins (PAs)- to develop a hemostatic device with improved properties. The effects of incorporating PAs into the aerogels were investigated in relation to their physicochemical properties, absorption ability, clotting activity and cytotoxicity in human dermal fibroblast (HDF) cells. The aerogels presented highly resistant porous structures, capable of absorbing more than 50 times their weight when in contact with a phosphate saline solution (PBS) and fresh human blood. Interestingly, the addition of PAs increased the negative surface charges and the blood absorption ability of the aerogels, which may make them suitable for hemostasis. The incorporation of 5% and 10% (w/w) of extracts into the aerogels increased the total coagulated blood content by 36.6% and 24.5% compared with gelatin-GO aerogel, respectively. These improvements in the hemostatic properties of the aerogels were greater with the inclusion of 5% (w/w) of grape skin extracts into the aerogels. The aerogels were also able to adhere red blood cells onto their surfaces, which could favor the formation of stable fibrin networks to promote hemostasis. Their clotting activity suggested the activation of alternative routes based on complement coagulation systems. Finally, the aerogels were non-toxic for HDF cells and the PAs were successfully released from their matrices. Thus, gelatin-GO aerogels reinforced with PAs are promising as topical phytodrug delivery systems, with great potential for wound healing processes., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Epileptic syndromes are well-defined conditions comprising particular clinical features [seizure types, age of onset, response to treatment] and characteristic electroencephalographic changes, while their etiology and subsequent prognosis may vary. The recognition of these syndromes is fundamental for pediatric neurology practice, representing an essential learning topic in this field. Nevertheless, many epileptic syndromes are still quite unfamiliar to students, residents and even neurologists, because of their low incidence and their minimal representation in the literature. This narrative review discusses the concept of epileptic syndromes and revisits seven lesser-known or uncommon syndromes in order to summarize their core clinical features, which can become important clues for daily neurological practice, namely epilepsy of infancy with migrating focal seizures, myoclonic epilepsy of infancy, self-limited infantile epilepsy, myoclonic encephalopathy in nonprogressive disorders, Jeavons syndrome, and epilepsy with myoclonic absences., (Copyright © 2021. Published by Elsevier Ltd.)

Introduction: Familial Hypercholesterolemia (FH) is an autosomal dominant disease with an estimated prevalence between 1/200-250. It is under-treated and underdiagnosed. Massive data screening can increase the detection of patients with FH., Methods: Study population: Residents in the health coverage area (N: 195.000 inhabitants) and with at least one determination of cholesterol linked to low-density lipoproteins (LDL-C) carried out between January 1, 2010 and December 30, 2019. The highest LDL-C values were selected., Exclusion Criteria: nephrotic syndrome, hypothyroidism, Hypothyroid treatment or triglycerides> 400 mg / dL. Seven algorithms suggestive of Familial Hypercholesterolemia Phenotype (HF-P) were analyzed, selecting the most efficient algorithm that could easily be translated into clinical practice., Results: Based on 6.264.877 assistances and 288.475 patients, after applying the inclusion-exclusion criteria, 504.316 tests were included, corresponding to 106.382 adults and 10.509 <18 years. The selected algorithm presented a prevalence of 0.62%. 840 patients with HF-P were detected, 55.8% being women and 178 <18 years old, 9.3% had a history of cardiovascular disease (CVD) and 16.4% had died. 65% of the patients in primary prevention had LDL-C values> 130 mg / dL and 83% in secondary prevention values> 70mg / dL. A ratio of 7.64 (1-18) patients with HF-P per analytical requesting physician was obtained., Conclusions: Massive data screening and patient profiling are effective tools and easily applicable in clinical practice for the detection of patients with FH., (Copyright © 2020 Sociedad Española de Arteriosclerosis. Publicado por Elsevier España, S.L.U. All rights reserved.)

The acquisition of neuroendocrine (NE) characteristics by prostate cancer (PC) cells relates to tumor progression and hormone resistance. PC cells may survive and function in androgen-deprived environments, where they could establish paracrine signaling networks, providing stimuli for the propagation of local carcinoma cells. We previously demonstrated, using electron probe X-ray microanalysis (EPXMA), in LNCaP, PC-3, and Du 145 cell lines that apoptosis is associated with intracellular elemental changes, and that the NE secretory products, bombesin and calcitonin, inhibit etoposide-induced apoptosis, as well as some of these elemental changes. In this study, LNCaP cells were induced in vitro to transdifferentiate under androgen deprivation, to mimic the role of NE cells in the apoptotic activity of transdifferentiated androgen-dependent PC cells. Changes in intracellular ion content associated with apoptosis, assessed by EPXMA, demonstrate that the transdifferentiated LNCaP cells are resistant to etoposide-induced apoptosis and also to the etoposide-induced elemental changes. The aggressive malignant potential of PC with neuroendocrine differentiation, associated with hormonal independence, is partly because of the ability that most NE tumor cells have to escape apoptosis, which can enhance the malignant properties of tumor cells and may have therapeutic implications as tumor cells are usually resistant to cytotoxic drugs as etoposide.

New biomaterials for Guided Bone Regeneration (GBR), both resorbable and non-resorbable, are being developed to stimulate bone tissue formation. Thus, the in vitro study of cell behavior towards material surface properties turns a prerequisite to assess both biocompatibility and bioactivity of any material intended to be used for clinical purposes. For this purpose, we have developed in vitro studies on normal human osteoblasts (HOB ® ) HOB ® osteoblasts grown on a resorbable Poly (lactide-co-glycolide) (PLGA) membrane foil functionalized by a very thin film (around 15 nm) of TiO 2 ( i.e. , TiO₂/PLGA membranes), designed to be used as barrier membrane. To avoid any alteration of the membranes, the titanium films were deposited at room temperature in one step by plasma enhanced chemical vapour deposition. Characterization of the functionalized membranes proved that the thin titanium layer completely covers the PLGA foils that remains practically unmodified in their interior after the deposition process and stands the standard sterilization protocols. Both morphological changes and cytoskeletal reorganization, together with the focal adhesion development observed in HOB osteoblasts, significantly related to TiO₂ treated PLGA in which the Ti deposition method described has revealed to be a valuable tool to increase bioactivity of PLGA membranes, by combining cell nanotopography cues with the incorporation of bioactive factors.

The possibility of tailoring membrane surfaces with osteoconductive potential, in particular in biodegradable devices, to create modified biomaterials that stimulate osteoblast response should make them more suitable for clinical use, hopefully enhancing bone regeneration. Bioactive inorganic materials, such as silica, have been suggested to improve the bioactivity of synthetic biopolymers. An in vitro study on HOB human osteoblasts was performed to assess biocompatibility and bioactivity of SiO2 functionalized poly(lactide-co-glycolide) (PLGA) membranes, prior to clinical use. A 15 nm SiO2 layer was deposited by plasma enhanced chemical vapour deposition (PECVD), onto a resorbable PLGA membrane. Samples were characterized by X-ray photoelectron spectroscopy, atomic force microscopy, scanning electron microscopy, and infrared spectroscopy (FT-IR). HOB cells were seeded on sterilized test surfaces where cell morphology, spreading, actin cytoskeletal organization, and focal adhesion expression were assessed. As proved by the FT-IR analysis of samples, the deposition by PECVD of the SiO2 onto the PLGA membrane did not alter the composition and other characteristics of the organic membrane. A temporal and spatial reorganization of cytoskeleton and focal adhesions and morphological changes in response to SiO2 nanolayer were identified in our model. The novedous SiO2 deposition method is compatible with the standard sterilization protocols and reveals as a valuable tool to increase bioactivity of resorbable PLGA membranes.

The specific adsorption of oxygenated and aliphatic gasoline components onto activated carbons (ACs) was studied under static and dynamic conditions. Ethanol and n-octane were selected as target molecules. A highly porous activated carbon (CA) was prepared by means of two processes: carbonization and chemical activation of olive stone residues. Different types of oxygenated groups, identified and quantified by TPD and XPS, were generated on the CA surface using an oxidation treatment with ammonium peroxydisulfate and then selectively removed by thermal treatments, as confirmed by TPD results. Chemical and porous transformations were carefully analyzed throughout these processes and related to their VOC removal performance. The analysis of the adsorption process under static conditions and the thermal desorption of VOCs enabled us to determine the total adsorption capacity and regeneration possibilities. Breakthrough curves obtained for the adsorption process carried out under dynamic conditions provided information about the mass transfer zone in each adsorption bed. While n-octane adsorption is mainly determined by the porosity of activated carbons, ethanol adsorption is related to their surface chemistry, and in particular is enhanced by the presence of carboxylic acid groups., (Copyright © 2013 Elsevier B.V. All rights reserved.)

We have studied the effect of the UV induced superhydrophilic wetting of TiO(2) thin films on the osteoblasts cell adhesion and cytoskeletal organization on its surface. To assess any effect of the photo-catalytic removal of adventitious carbon as a factor for the enhancement of the osteoblast development, 100 nm amorphous TiO(2) thin layers were deposited on polyethylene terephthalate (PET), a substrate well known for its poor adhesion and limited wettability and biocompatibility. The TiO(2) /PET materials were characterized by X-ray photoelectron spectroscopy, and atomic force microscopy and their wetting behavior under light illumination studied by the sessile drop method. The amorphous TiO(2) thin films showed a very poor photo-catalytic activity even if becoming superhydrophilic after illumination. The illuminated samples recovered partially its initial hydrophobic state only after their storage in the dark for more than 20 days. Osteoblasts (HOB) were seeded both on bare PET and on TiO(2) /PET samples immediately after illumination and also after four weeks storage in darkness. Cell attachment was much more efficient on the immediately illuminated TiO(2)/PET samples, with development of focal adhesions and cell traction forces. Although we cannot completely discard some photo-catalytic carbon removal as a factor contributing to this cell enhanced attachment, our photodegradation experiments on amorphous TiO(2) are conclusive to dismiss this effect as the major cause for this behavior., (Copyright © 2012 Wiley Periodicals, Inc.)

Osseointegration of implants is crucial for the long-term success of oral implants. The periimplant bone formation by osteoblasts is strongly dependent on the local mechanical environment in the interface zone. Robust demands for energy are placed on osteoblasts during the adhesion process to solid surfaces, and mitochondria are capital organelles in the production of most of the ATP needed for the process. We have assessed the relationship between osteoblast differentiation and mitochondrial bioenergetics in living cells grown on two different titanium surfaces, in order to provide valuable information for the design of material surfaces required for the development of the most appropriate osteogenic surface for osteoblastic anchorage. Combined backscattered and fluorescence confocal microscopy showed that in flat cells grown on a machined surface, highly energized mitochondria were distributed along the cell body. In contrast, cells grown on the rough surface emitted long protrusions in search of surface roughness, with actin stress fibers clearly polarized and highly energized mitochondria clustered at focal adhesion sites. This report using normal human osteoblastic cells indicates that these cells are especially sensitive to surface cues through energy production that enhances the necessary adhesion required for a successful osseointegration.

To dissect the rat receptor for advanced glycation end products (RAGE) subcellular distribution and trafficking in eukaryotic cells, an expression system coding for a fusion protein between the RAGE and an enhanced green fluorescent protein (EGFP) has been used. The RAGE-EGFP protein is expressed at the plasma membrane of CHO-k1 and Neuro-2a (N2a) cells and retains the capacity to bind Texas Red-labelled advanced glycation end products (AGEs). AGEs addition to the cell cultures induced a change in the subcellular distribution of the fluorescent RAGE-EGFP protein compatible with an internalization of the AGEs-RAGE complex. Furthermore, while N2a cells expressing the RAGE-EGFP showed an increase in ERK1/2 phosphorylation and NF-kappaB DNA binding in response to AGEs, pre-incubation with dansyl-cadaverine or phenylarsine oxide, inhibitors of receptors internalization, blocked the activation of ERKs and other intracellular responses mediated by AGEs. These results suggest that internalization plays a key role in the signal transduction mediated by RAGE.

The understanding of the cellular basis of osteoblastic cell-biomaterial interaction is crucial to the analysis of the mechanism of osseointegration. Cell adhesion is a complex process that is dependent on the cell types and on the surface microtopography and chemistry of the substrate. We have studied the role of microtopography in modulating cell adhesion, in vitro, using a human osteoblastic cell line for the assessment of actin cytoskeletal organization. Through application of CLSM combining reflection and fluorescence, 2D or 3D images of cytoskeleton were obtained. On smooth surfaces, Ti CP machined, predominantly planar bone cells with an axial ratio of 1.1 were randomly oriented, with stress fibers running in all directions, and thin filopodia. On TiCP Osseotite surfaces the osteoblastic cells conformed to the irregular terrain of the sustrate with focal adhesion sites only established on the relative topographical peaks separated for a longer distance than in the machined surface, and defined wide lamellopodia and long filopodia, with enhanced expression of stress fibers, forming large clear focal contacts with the rough surface. The cytoskeletal organization of cells cultured on rough titanium supports an active role for the biomaterial surface in the events that govern osteoblastic cell adhesion. The results enforce the role of the rough sustrate surface in affecting osteoblastic cell adhesion and provide valuable information for the design of material surfaces that are required for the development of an appropriate osteogenic surface for osteoblastic anchorage, compared to machined surface, in dental implants.

Neuroendocrine secretory products and their interactions with epithelial prostate cells are currently under investigation in order to understand their significance in the pathogenesis, prognosis, and therapy of prostate carcinoma. These neuropeptides have the potential to disrupt the balance between cell death and cell growth in the tumor. Our research was based on the role of bombesin in modulating the mitochondrial membrane potential (Delta psi(m)) in cell death induced by etoposide on PC-3 cells. Cells were cultured and stained with 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine iodide (JC-1). At low membrane potentials, JC-1 produces a green fluorescence, and at high membrane potentials, it forms "J aggregates" with red fluorescence. Cells were examined in a confocal microscope. For quantitative analyses, regions of interest were selected. The size, number of pixels, and ratios between fluorescence intensity in the red and green channels in each region of interest were calculated. The loss of Delta psi(m) in etoposide-treated PC-3 cells was prevented by bombesin. The quantitative analysis of JC-1-stained cells revealed a significant decrease in the red (high Delta psi(m)) to green (low Delta psi(m)) ratio in etoposide-treated cells when compared with control cells, which was restored in the presence of bombesin (P < 0.00001). The interaction between treatments and area (P = 0.0002) was highly significant, and confirms that PC-3 cells keep their apoptosis machinery, showing an apoptotic volume decrease in response to etoposide. The protection by bombesin occurs by inhibition of apoptosis and maintenance of mitochondrial integrity. New therapeutic protocols and trials need to be developed to test drugs acting through the neutralization of antiapoptotic intracellular pathways mediated by neuroendocrine hormones.

It has been suggested that neuroendocrine (NE) cells provide paracrine stimuli for the propagation of local carcinoma cells and that NE differentiation is associated with the progression of prostate cancer toward an androgen-independent state. Apoptosis comprises a critical intracellular defense mechanism against tumorigenic growth and is associated with a number of changes in the elemental content of the cell. The neuropeptides bombesin and calcitonin, which inhibit etoposide-induced apoptosis, also inhibit the etoposide-induced elemental changes in prostate carcinoma cells. This important fact strengthens the link between apoptosis and changes in the intracellular elemental content. This protective effect on etoposide-induced apoptosis appears to be quite similar in androgen-dependent and androgen-independent cell lines. This confirms that neuropeptides confer antiapoptotic capabilities on non-neuroendocrine cells in close proximity to neuroendocrine cells. It can therefore be speculated that certain neuroendocrine peptides can increase the survival and further growth of neighboring cells and may thereby contribute to the aggressive clinical course of prostate tumors containing neuroendocrine elements. In addition, this correlation provides an objective basis for the study of neuropeptide target points and may be helpful for alternative therapeutic protocols using neuropeptide inhibitors in the treatment of patients with advanced prostatic carcinoma. The culture techniques described were, thus, designed in order to achieve two important goals. First, the development of an in vitro model that allows an approach to neuroendocrine differentiation in prostate cancer and its role in apoptosis blockage. Second, the method has been designed in order to permit rapid cryofixation of intact cell monolayers for subsequent x-ray microanalysis.

Eosinophilic gastroenteritis (EGE) is a rare inflammatory disease characterized by diffuse or scattered eosinophilic infiltration of the digestive tract and usually by peripheral blood eosinophilia. The most common presenting symptoms of EGE are abdominal pain, vomiting and diarrhea, but clinical features depend on which layers or location of gastrointestinal tract are involved. Treatment with corticosteroids results in clinical and histological remission in most patients and surgery can be avoided if a correct diagnosis is made. Previous history of allergy is a key to diagnosing EGE, but peripheral eosinophilia may be absent in some patients under concomitant treatment with corticosteroids. Radiological and endoscopic findings are also nonspecific and diagnosis must always be histologically confirmed. The gastrointestinal involvement is patchy in distribution, so more than one panendoscopic examination is often necessary to establish the diagnosis, and surgical or CT-guided full-thickness biopsy is needed in patients with muscular or serosal involvement. It emphasises the importance of a high index of clinical suspicion, which mainly depends on knowledge of natural history of the disease. We report here a case of EGE associated with transmural eosinophilic cholecystocholangitis, in a patient who presented with dyspeptic symptoms and recurrent cholestasis responsive to corticoesteroids. To our knowledge, this patient represents the second case, in the English literature, in which corticoid-responsive cholangitis was associated to histologically proven eosinophilic cholecystitis and gastrointestinal involvement, suggesting that EGE must always be considered in the differential diagnosis of biliary tract disease in patients with eosinophilia and/or atopic diseases.

Background: Etoposide-induced apoptosis in prostate carcinoma cells is associated with changes in the elemental content of the cells. The authors previously reported that calcitonin and bombesin inhibited etoposide-induced apoptosis in these cells. In the current study, the authors investigated whether these neuropeptides block the etoposide-induced changes in elemental content., Methods: Cells from the PC-3 and Du 145 prostate carcinoma cell lines were grown either on solid substrates or on thin plastic films on titanium electron microscopy grids, and they were exposed to etoposide for 48 hours in the absence or presence of calcitonin and bombesin. After the exposure, the cells were frozen and freeze dried, and their elemental content was analyzed by energy-dispersive X-ray microanalysis in both in the scanning electron microscope and the scanning transmission electron microscope., Results: Etoposide treatment consistently induced an increase in the cellular Na concentration and a decrease in the cellular K concentration, resulting in a marked increase of the Na/K ratio and also an increase in the phosphorus:sulphur (P/S) ratio. Both bombesin and calcitonin inhibited the etoposide-induced changes in the cellular Na/K ratio, and calcitonin, but not bombesin, inhibited the changes in the P/S ratio. No significant elemental changes were found with bombesin or calcitonin alone., Conclusions: The neuropeptides bombesin and calcitonin, which inhibited etoposide-induced apoptosis, also inhibited the etoposide-induced elemental changes in prostate carcinoma cells. This important fact strengthens the link between apoptosis and changes in the intracellular elemental content. This correlation provides an objective basis for the study of neuropeptide target points and may be helpful for alternative therapeutic protocols using neuropeptide inhibitors in the treatment of patients with advanced prostatic carcinoma.

Aims: We tried to show the demographic characteristic and alcohol intake habits among our outpatients. We study the influence of age, sex, habitat and socioeconomical status on alcoholic habit., Design: Retrospective and institution based study. Patients. 164 patients who were followed up for alcohol liver disease in our outpatient section., Results: Average age to start drinking alcohol was 18.6 (7.36) years, years of alcoholism were 35.4 (13.5) years, average daily alcohol intake was 161.2 (116.7) grams of pure alcohol. Only 16 men (8%) drank less than 60 grams a day. 5 (35.7%) women drank less than 40 grams a day. Life-cumulative alcohol intake was correlated with Maddrey's score at the end of the study (r = +0.407). Average daily alcohol intake was correlated with ultrasonographic features of the liver (r = +0.283), we appreciated that Prothrombin Time was also correlated with ultrasonographic features of the liver (r = +0.301). The percentage of patients who suffer, at least one decompensation of their disease was 39%., Conclusions: Average age to start drinking is about legal age. Life-cumulative alcohol intake was related to Prothrombin Time and ultrasonographic features of the liver.

Aging deeply influences several morphologic and functional features of the peripheral nervous system (PNS). Morphologic studies have reported a loss of myelinated and unmyelinated nerve fibers in elderly subjects, and several abnormalities involving myelinated fibers, such as demyelination, remyelination and myelin balloon figures. The deterioration of myelin sheaths during aging may be due to a decrease in the expression of the major myelin proteins (P0, PMP22, MBP). Axonal atrophy, frequently seen in aged nerves, may be explained by a reduction in the expression and axonal transport of cytoskeletal proteins in the peripheral nerve. Aging also affects functional and electrophysiologic properties of the PNS, including a decline in nerve conduction velocity, muscle strength, sensory discrimination, autonomic responses, and endoneurial blood flow. The age-related decline in nerve regeneration after injury may be attributed to changes in neuronal, axonal, Schwann cell and macrophage responses. After injury, Wallerian degeneration is delayed in aged animals, with myelin remnants accumulated in the macrophages being larger than in young animals. The interaction between Schwann cells and regenerative axons takes longer, and the amount of trophic and tropic factors secreted by reactive Schwann cells and target organs are lower in older subjects than they are in younger subjects. The rate of axonal regeneration becomes slower and the density of regenerating axons decrease in aged animals. Aging also determines a reduction in terminal and collateral sprouting of regenerated fibers, further limiting the capabilities for target reinnervation and functional restitution. These age-related changes are not linearly progressive with age; the capabilities for axonal regeneration and reinnervation are maintained throughout life, but tend to be delayed and less effective with aging.

Objective: To evaluate the utility of carbohydrate deficient transferrin (CDT) for the chronic alcoholism diagnostic, the recent intake and the abstinence, comparing it with the most recognized biological markers, specially the MCV and the GGT., Methods: We study 87 subjects: 22 controls, 42 subjects abstinent chronic alcoholics and 23 patient active alcoholics. From the last group we subject the middle to a 15 days abstinence period. We compare the CDT with the mos recognized markers in the difference groups., Results: We found significant difference with MCV and GGT in the chronic alcoholism diagnostic. There were significant difference with MCV, GGT and specially CDT in the case of recent alcoholic intake. CDT was the only one that shows significant difference in recent abstinence., Conclusion: According to our results the CDT is a poor marker of chronic alcoholism. The classical markers (MCV nd GGT) are better on tha propriety. CDT, however, is a valid and effective marker for the recent alcohol intake, it is better than GGT and MCV. And it is the only valid marker for the recent abstinence. So, we estimate that, it would be advisable the CDT clinical use with restringed criterions for the recent alcoholic intake diagnostics and, specially for the abstinence one.

Background: Neuroendocrine differentiation in prostatic carcinoma has been related to regulation of proliferation and metastatic potential and correlated with prognosis. More than 80% of prostate carcinomas initially respond to androgen ablation, but most relapse, due to the heterogeneous presence of androgen-dependent and independent clones. The pathways of cellular proliferation and apoptosis are inexorably linked to minimize the occurrence of neoplasia, and disfunction of apoptosis is proposed as a pathogenic process in malignant tumors. Androgen-dependent prostatic cancer cells undergo apoptosis after androgen deprivation, but not androgen-independent ones due to a defect in the initiation step. Anyway, they retain the basic cellular machinery to undergo apoptosis. We suggest a possible role of neuroendocrine differentiation in the onset and regulation of apoptosis in prostatic neoplasia., Methods: LNCaP, PC-3 and DU 145 prostatic cancer cell lines were induced to undergo apoptosis after treatment with etoposide alone or plus androgen ablation. We tested the role of neuropeptides bombesin and calcitonin at modulating etoposide induced apoptosis., Results: Etoposide-induced apoptosis in all cancer cell lines was achieved. In LNCaP androgen ablation was also required. Apoptosis is prevented in all three lines when bombesin was added. Calcitonin addition prevents apoptosis in PC-3, LNCaP and in an etoposide dose-dependent way in DU 145., Conclusion: Neuropeptides bombesin and calcitonin can modulate the apoptotic response of prostate cancer cells by inducing resistance to etoposide-induced apoptosis, suggesting that neuropeptides can be used as a target of therapeutical approach in prostatic carcinoma.

The nicotinic axon reflex-mediated sudomotor response was studied in mice and rats by recording the impressions of sweat droplets made in silicone molds after local injection of nicotine, and compared with sweating induced by acetylcholine and pilocarpine. Nicotine failed to activate mouse plantar sweat glands at any of the concentrations used (from 3 x 10(-6) to 3 x 10(-1) M). On the contrary, both acetylcholine and pilocarpine produced a dose-dependent increase in the number of secreting sweat glands. The location of sweat glands reactive to pilocarpine and acetylcholine was similar and restricted to the pads near the site of injection. We conclude that the sudomotor axon reflex response mediated by nicotinic receptors is not present in the mouse and the rat.

Metastatic prostatic cancer is typically refractory to androgen ablation therapy due to the presence of androgen-independent clones in the neoplasia. A therapeutical approach which could effectively control androgen-dependent and independent cells is, thus, needed. Maybe the failure of certain cancer cells to engage in apoptosis could explain the inherent drug resistance of many tumors. Anyway, these cells can retain the ability to undergo apoptosis in response to an adequate stimulus. We tested whether etoposide, a topoisomerase II inhibitor, could induce apoptosis in androgen-dependent (LNCaP) as well as independent (PC-3 and DU 145) human prostate cancer cell lines. Morphological examination was performed, as it is regarded as one of the most reliable parameters for the detection of apoptotic changes. Complementarily, biochemical and flow cytometric studies were also used. Characteristical changes of apoptosis were demonstrated in PC-3, Du 145, and LNCaP cancer cells after treatment with etoposide. These cells, thus, retain the ability to undergo apoptosis under adequate conditions, in a promising approach to hormone refractory prostate cancer therapy.

Functional sudomotor responses have been studied in sweat glands reinnervated after sciatic nerve crush and partially denervated by cisplatin intoxication in the mouse. The sudomotor function mediated by the sciatic nerve was evaluated by silicone imprints on the plantar surface of the hindpaws. Five days after nerve crush, completely denervated sweat glands became unresponsive to cholinergic stimulation with pilocarpine. During the following weeks, the number of reinnervated, reactive sweat glands increased progressively to reach a maximum of 89% of preoperative control counts by 40 days after nerve crush. At this time, the mean volume of sweat secreted per gland was normal, but reinnervated glands showed a secretory activity abnormally sustained over time after pilocarpine stimulation and, on the other hand, had an increased resistance to the inhibition of secretion induced by atropine. The effects of cisplatin administration on sudomotor function were investigated in two groups of mice, one treated with high doses of cisplatin (10 mg/kg/week for 4 weeks) and another treated with low doses of cisplatin (5 mg/kg/week for 8 weeks). Cisplatin intoxication produced abnormal sudomotor responses indicative of denervation from cumulative doses of 10 mg/kg. The first abnormality found was a partial resistance of sweat glands to atropine, followed by a decrease in the sweat output per gland and finally a decline in the number of sweat glands activated by pilocarpine. These abnormalities in the sudomotor responses were more pronounced in mice treated with a high dose than in those with a lower dose regime.