A universal dose-response curve for radiochromic films.

PURPOSE: This paper presents a model for dose-response curves of radiochromic films. It is based on a modified version of single-hit model to take into account the growth experienced by lithium salt of pentacosa-10,12-diynoic acid polymers after irradiation. METHODS: Polymer growth in radiochromic films is a critical phenomenon that can be properly described by means of percolation theory to provide an appropriate distribution function for polymer sizes. Resulting functional form is a power function featuring a critical exponent and two adjustable parameters. Moreover, these parameters act as scaling factors setting a natural scale for sensitometric curves where the dependence on channel sensitivity is removed. A unique reduced response curve is then obtained from all the color channels describing film behavior independently of film dosimetry system. RESULTS: Resulting functional form has been successfully tested in several sensitometric curves from different Gafchromic EBT models, providing excellent agreement with experimental data in a wide dose range up to about 40 Gy and low dose uncertainty. CONCLUSIONS: The model presented in this paper describes accurately the sensitometric curves of radiochromic films in wide dose ranges covering all typical ranges used in external radiotherapy. Resulting dose uncertainty is low enough to render a reasonably good performance in clinical applications. Due to cross-correlation, only one of the adjustable parameters is totally independent and characterizes film batches.

Molecular characterization of calcitonin gene-related peptide (CGRP) related peptides (CGRP, amylin, adrenomedullin and adrenomedullin-2/intermedin) in goldfish (Carassius auratus): cloning and distribution.

To further characterize the structure and function of calcitonin gene-related peptide (CGRP) related peptides in fish, we have cloned cDNA sequences for CGRP, amylin, adrenomedullin (AM) and adrenomedullin-2/intermedin (IMD) in goldfish (Carassius auratus) and examined their tissue distribution. CGRP, amylin, AM and IMD cDNAs were isolated by reverse transcription (RT) and rapid amplification of cDNA ends (RACE). The cloned sequences contain the complete four mature peptides, which present a high degree of identity with mature peptide sequences from other fish. Phylogenetic analyses show that goldfish AM and IMD form a sub-family within the CGRP-related peptides that is distinct from the CGRP/amylin sub-family. The distribution of goldfish CGRP-like peptides mRNA expression in different tissues and within the brain was studied by RT-PCR. CGRP, IMD and AM are detected throughout the brain, in pituitary and in most peripheral tissues examined. Amylin mRNA is mostly expressed in the brain, in particular posterior brain, optic tectum and hypothalamus, but is also present in pituitary, gonad, kidney and muscle. Our results suggest that goldfish CGRP, amylin, AM and IMD are conserved peptides that show the typical structure characteristics present in their mammalian counterparts. The widespread distributions of CGRP, AM and IMD suggest that these peptides could be involved in the regulation of many diverse physiological functions in fish. Amylin mRNA distribution suggests possible new roles for this peptide in teleosts, including the control of reproduction.

Expression of gonadotrophin-releasing hormone binding sites in somatic tissues of the gilthead seabream (Sparus aurata): a quantitative autoradiographic study.

In this study, we have analysed the expression of gonadotrophin-releasing hormone (GnRH) binding sites in somatic tissues (intestine, liver, gill, skeletal muscle, ovary, heart, stomach, kidney and spleen) of the gilthead seabream, Sparus aurata using 3-[125I]iodototyrosyl5-mammalian GnRH and auto-radiographic techniques. The qualitative and quantitative analysis showed the existence of a basal expression of specific GnRH binding sites in intestine, skeletal muscle, ovary, stomach and spleen. Furthermore, our data suggest that the level of expression of GnRH binding sites can be significantly enhanced by GnRH treatment in intestine, gill, heart, stomach, kidney and spleen. This study shows that GnRH can exert direct effects in both reproductive and non-reproductive somatic tissues of the gilthead seabream.

Morbidity of laparoscopic surgery for complicated appendicitis: an international study.

BACKGROUND: Although laparoscopic appendectomy has some advantages over open appendectomy, some reports do show more postoperative intraabdominal abscesses. METHODS: A retrospective review of complicated appendicitis managed surgically by eight surgical groups from six countries was undertaken. Among 3,433 patients with appendicitis, 1,017 (29.5%) had complicated appendicitis, which included perforated or gangrenous appendicitis with or without localized or disseminated peritonitis. There were 74 preoperative abscesses (7.4%) and 5 small bowel obstructions. RESULTS: One patient died. There were 29 postoperative intraabdominal abscesses (2.8%) and 112 mostly minor complications. Conversion to laparotomy was necessary for 28 patients (2.7%). The surgical time ranged from 32 to 132 min (mean, 62 min), and the hospital stay ranged from 1 to 18 days (mean, 3.5 days). CONCLUSIONS: The morbidity rates, particularly for intraabdominal abscesses, were less for laparoscopic appendectomy in complicated appendicitis than those reported in the literature for open appendectomy, whereas operating times and hospital stays were similar.

The GnRH system in the European sea bass (Dicentrarchus labrax).

The cDNA sequences encoding three GnRH forms, sea bream GnRH (sbGnRH), salmon GnRH (sGnRH) and chicken GnRH II (cGnRH II), were cloned from the brain of European sea bass, Dicentrarchus labrax. Comparison of their deduced amino acid sequences to the same forms in the gilthead sea bream, Sparus aurata, and striped bass, Morone saxatilis, revealed high homology of the prepro-cGnRH II (94% and 98% respectively), and prepro-sGnRH (92% to both species). The sbGnRH exhibited dissimilar identities, with high homology to the striped bass (93%), and lower homology (59%) to the gilthead sea bream. Two transcript types were identified for the GnRH-associated peptide (GAP)-sGnRH as well as for the GAP-cGnRH II, which suggests a possible alternative splicing followed by the addition of an early stop codon. In order to obtain antibodies specific for the three GnRH precursors, recombinant GAP proteins were produced. The differential expression of the three GnRHs previously reported in the brain by means of in situ hybridization, using riboprobes corresponding to the GAP-coding regions, was fully confirmed by immunocytochemistry using antibodies raised against the recombinant GAP proteins, indicating that the transcripts are translated into functional proteins. Moreover, this approach allowed us to follow, for the first time, the specific projections of the different cell groups: sGAP fibers are distributed mainly in the forebrain with few projections reaching the pituitary, sbGAP fibers are mainly present in the preoptic area, mediobasal hypothalamus and predominantly project to the pars distalis of the pituitary, whereas cGnRH II fibers have a widespread distribution primarily in the posterior brain, and do not project to the pituitary. These new tools will be extremely useful to study further the development, regulation and functional significance of three independent GnRH systems in the brain of vertebrate species.

Laparoscopic repair of traumatic diaphragmatic hernia.

Posttraumatic diaphragmatic rupture or eventration is still a challenging problem. Herein five cases of patients with such a diaphragmatic lesion treated successfully by laparoscopy are reported with a discussion of the advantages of this mini-invasive surgical approach.

Immunocytohistochemical characterization of pituitary cells of the bluefin tuna, Thunnus thynnus L.

In this paper we report the first complete mapping of the pituitary in a tuna species. The various different adenohypophysis cell types of the bluefin tuna, Thunnus thynnus L. have been identified and located using different antisera against mammalian and piscine hormones and various histochemical techniques: PAS, Alcian Blue pH 2.5 and lectins -ConA and WGA(Neutral and Acidic Glycoproteins); Bromophenol Blue (Proteins) and Tioglycollate-Ferric-Ferricianide-FeIII (-S-S- groups). Prolactin (PRL) and adrenocorticotrophic (ACTH) cells were located in the rostral pars distalis (RPD) of the pituitary, while the proximal pars distalis (PPD) displayed gonadotrophic (GTH), thyrotrophic (TSH), somatotrophic (GH) and also a few PRL cells. Moreover, somatolactin (SL) and melanotrophic (MSH) cells were identified inside the pars intermedia (PI). Interestingly, some SL-immunoreactive fibers were also detected in the neurohypophysis. Some GTH cells were also located on the exterior surface of the PI. Glycoproteins containing mannose (Man) and/or glucose (Glc); N-acetyl-glucosamine (GlcNAc) and/or sialic acid sugar residues, as well as -S-S- groups, were observed in GTH, TSH and SL cells. The Bromophenol Blue technique stained amphiphilic SL, acidophilic GH cells and weakly ACTH cells. GH and ACTH cells were unreactive to PAS, Alcian Blue, Tioglycollate-Ferric-Ferricianide-FeIII and lectin (Con A and WGA) techniques. Finally, PAS reaction was positive in amphiphilic SL cells, which were PbH unreactive, while MSH and ACTH cells were stained with PbH technique.

Distribution of neuropeptide Y-like immunoreactivity in the brain of the Senegalese sole (Solea senegalensis).

We present the results of an immunohistochemical study aimed at localizing the neuropeptide Y (NPY) in the brain of the Senegalese sole, Solea senegalensis, using an antiserum raised against porcine NPY and the streptavidin-biotin-peroxidase method. In this species, we have identified immunoreactive cells in the ventral and dorsal telencephalon, caudal preoptic area, ventrocaudal hypothalamus, optic tectum, torus longitudinalis, synencephalon and isthmic region. NPY-immunoreactive fibers were profusely distributed throughout the brain, also reaching the adenohypophysis. The extensive distribution of NPY suggests an important role for this neuropeptide in a variety of physiological processes, including the neuroendocrine control of adenohypophyseal functions. Our results are compared with those obtained in other teleosts and discussed in relation to putative functions of NPY in the control of metabolism and reproduction in the Senegalese sole.

Differential expression of three different prepro-GnRH (gonadotrophin-releasing hormone) messengers in the brain of the european sea bass (Dicentrarchus labrax).

The expression sites of three prepro-gonadotrophin-releasing hormones (GnRHs), corresponding to seabream GnRH (sbGnRH: Ser(8)-mGnRH, mammalian GnRH), salmon GnRH (sGnRH: Trp(7)Leu(8)-mGnRH), and chicken GnRH-II (cGnRH-II: His(5)Trp(7)Tyr(8)-mGnRH) forms were studied in the brain of a perciform fish, the European sea bass (Dicentrarchus labrax) by means of in situ hybridization. The riboprobes used in this study correspond to the three GnRH-associated peptide (GAP)-coding regions of the prepro-GnRH cDNAs cloned from the same species (salmon GAP: sGAP; seabream GAP: sbGAP; chicken GAP-II: cIIGAP), which show little oligonucleotide sequence identity (sGAP versus sbGAP: 42%; cIIGAP versus sbGAP: 36%; sGAP versus cIIGAP: 41%). Adjacent paraffin sections (6 mm) throughout the entire brain were treated in parallel with each of the three anti-sense probes and the corresponding sense probes, demonstrating the high specificity of the hybridization signal. The results showed that both sGAP and sbGAP mRNAs had a broader expression in the olfactory bulbs, ventral telencephalon, and preoptic region, whereas cIIGAP mRNA expression was confined to large cells of the nucleus of the medial longitudinal fascicle. In the olfactory bulbs, both the signal intensity and the number of positive cells were higher with the sGAP probe, whereas sbGAP mRNA-expressing cells were more numerous and intensely stained in the preoptic region. Additional isolated sbGAP-positive cells were detected in the ventrolateral hypothalamus. These results demonstrate a clear overlapping of sGAP- and sbGAP-expressing cells in the forebrain of the European sea bass, in contrast to previous reports in other perciforms showing a clear segregation of these two cell populations.

Laparoscopic diagnosis and treatment of an acute epiploic appendagitis with torsion and necrosis causing an acute abdomen.

Acute epiploic appendagitis is not as rare as previously thought; but, since the presenting signs and symptoms are not specific, preoperative diagnosis has been rarely made. At the present time, a laparoscopic exploration of the peritoneal cavity will establish the correct diagnosis, and the treatment can be provided during the same procedure. Herein, a case of a 63-year-old female patient with acute abdominal syndrome caused by a necrotic epiploic appendix that was successfully diagnosed and treated laparoscopically is described. A review of the literature is made, as well.

Technical modifications for laparoscopic appendetomy and other pelvic procedures using microinstruments.

Reusable microinstruments are used in a modification of the three-port technique for laparoscopic appendectomy and other pelvic procedures; the method is described herein. Fifty-seven consecutive patients were operated upon using this technique. There was no mortality; there were two minor complications and no conversions to laparotomy. Pain medication needs were minimal in all patients, and the cosmetic results were excellent. The use of microinstruments for pelvic laparoscopic procedures is safe and cost-effective using the technical modifications described.

Localization of tyrosine hydroxylase-immunoreactivity in the brain of the Senegalese sole, Solea senegalensis.

The localization of catecholamines in the brain of the Senegalese sole was determined by immunohistochemical techniques using antibodies against tyrosine hydroxylase. Although the general pattern of distribution of catecholamines is consistent with that reported in other teleosts, some remarkable differences are observed. The most rostral tyrosine hydroxylase immunoreactive (TH-ir) cells were identified in the olfactory bulbs, in which a clear asymmetry in the number and location of TH-ir perikarya and fibers was observed. The number of TH-ir cells is manifestly higher in the right olfactory bulb, especially in the internal cell layer. TH-ir fibers are also much more abundant in the right bulb, principally in the glomerular and internal cell layers. Other TH-ir cell masses were identified in the ventral telencephalon, preoptic area, caudoventral hypothalamus, posterior tuberculum, synencephalon, isthmic region and rhombencephalon. Surprisingly, no ir cell bodies were identified in the ventromedial thalamic nucleus, which exhibits a large number of TH-ir cells in other teleosts. The presence of TH-ir fibers in the brain of sole is particularly evident within and around the nuclei in which immunoreactive cells are found. However, other zones such as the dorsal telencephalon, posterior commissure, optic tectum, torus semicircularis, reticular formation or inferior olive also displayed TH-ir fibers. TH-ir axons also enter the infundibulum, reaching the proximal pars distalis of the adenohypophysis. The distribution of TH-ir cells and fibers is compared with that observed in other teleosts and is discussed in a comparative context.

Localization of galanin-like immunoreactive structures in the brain of the Senegalese sole, Solea senegalensis.

The distribution of galanin-like immunoreactive structures was studied in the brain of the Senegalese sole, Solea senegalensis, using immunohistochemical methods. Periventricular immunoreactive cell bodies were observed in the rostral pole of the preoptic recess, within the pars parvocellularis of the nucleus preopticus parvocellularis. Another galanin-immunoreactive cell population was observed more caudal in the ventromedial hypothalamus, along the medial evaginations of the lateral recess. These cells appear within the cytoarchitectonic limits of the nucleus recessus lateralis pars ventralis. We found an extensive presence of galanin-immunoreactive fibres throughout the entire brain, although the most massive network of fibres was observed in the caudal olfactory bulbs, ventral telencephalon, preoptic area and around diencephalic ventricular recesses. Also, the hypophysis, ventricular mesencephalic area, median reticular formation and viscerosensory rhombencephalon displayed important plexuses of galanin-immunoreactive axons. The widespread distribution of these immunoreactive structures in the brain and pituitary of the Senegalese sole suggests an important role for galanin in neuroendocrine regulation of brain and adenohypophyseal functions.

Distribution of serotonin in the brain of the Senegalese sole, Solea senegalensis: an immunohistochemical study.

We report the distribution of serotonin immunoreactive (5-HT-ir) structures in the brain of the adult Senegalese sole, Solea senegalensis, using the streptavidin-biotin-peroxidase complex immunohistochemical method. We have found a wide distribution of immunoreactive fibers throughout the entire brain. 5-HT-ir cell bodies appeared restricted to some periventricular nuclei associated with the diencephalic recesses, and in the rhombencephalic reticular formation and inferior olivary region. Specifically, cerebrospinal fluid-contacting serotoninergic cells were found within the pars dorsalis and pars ventralis of the nucleus recessus lateralis, in the paraventricular organ and in the nucleus recessus posterioris. In the brainstem, 5-HT-ir perikarya appear within the superior and inferior raphe, the nucleus reticularis superioris, the nucleus interpeduncularis and the inferior olive. Although positive fibers were not found in the neurohypophysis, a few 5-HT-ir cells were identified in the adenohypophysis. This distribution is compared with those found in other fishes and discussed in the context of putative roles of 5-HT as a neuroendocrine factor and neurotransmitter in the Senegalese sole.

Laparoscopic-assisted resection of a bleeding gastrointestinal stromal tumor.

The authors report a case of a 29-year-old male patient with a severe lower gastrointestinal hemorrhage in whom a successful laparoscopic diagnosis and resection (assisted) of an ileal gastrointestinal stromal tumor (GIST) was performed. Laparoscopy can be very useful in the diagnosis and treatment of selected cases of lower gastrointestinal bleeding.

[Critical vision in the treatment of invasive vesical cancer with ionizing radiation]. / Una visión crítica del tratamiento del cáncer vesical invasor con radiación ionizante.

OBJECTIVE: To review the treatment of invasive bladder cancer with ionizing radiation. METHODS: Patient selection criteria, prognostic factors, treatment of different tumor types with radiation using different strategies alone or in combination with chemotherapy are analyzed in the literature. RESULTS/CONCLUSIONS: The role of radiotherapy in the treatment of invasive carcinoma of the bladder is crucial: 1) preoperative radiotherapy in combination with radical surgery achieves results comparable with those of cystectomy in terms of survival and a higher local control rate; 2) used in combination with cisplatin post-TUR, irradiation is highly effective for local control of the tumor and bladder preservation, with complete remission rates ranging from 60-70%, which are higher than those of radical cystectomy; 3) in this latter therapeutic modality the effect of radiotherapy is dose-related and is influenced by factors such as tumor size, presence or absence of urinary obstruction, presence of multiple lesions on the bladder wall and extravesical spread. Over the last few years, a number of therapeutic procedures have been developed, which can improve the previously mentioned results when used appropriately. Among these are hyperfractionation, accelerated irradiation and concomitant radio and chemotherapy. Studies to determine tumor radiosensitivity (Fs2/a) and the use of gene therapy could enhance the tumor control rate in the future, although a system for the selection on patients for conservative or multilating treatment based on clinical, clinicopathological and scientific criteria will have to be developed. Finally, the use of special techniques, particularly brachytherapy and intraoperative irradiation, in combination with external radiotherapy has achieved excellent results (high local control and survival rates) in certain situations, basically in single/small-sized tumours.

Cloning and sequencing of the gilthead sea bream estrogen receptor cDNA.

We report here the complete nucleotide sequence of a cDNA clone containing the full-coding sequence of the Sparus aurata estrogen receptor (ER) isolated from an expression library prepared from gilthead sea bream liver poly A+ RNA. The library was screened using a single strand rainbow trout ER cDNA probe, corresponding to the C-D domain. The cDNA sequence containing an insert of 2369 nucleotides was found to encode a protein of 579 amino acids. The 5'- and 3'-untranslated regions of the message are 186 and 392 nucleotides long, respectively. The gilthead sea bream ER shows the higher homology with the ER of another perciform, Chrysophrys major (93%), moderate to high homology with Oreocromis aureus (78%) medaka (77%) and rainbow trout (70.7%) ERs and lower homology with japanese eel (45%), amphibian (47%), avian (48.5%) and mammalian (47-47.5%) ERs. The sequence homologies and phylogenetic analysis of the various ERs suggest that gilthead sea bream ER should be considered as a ER alpha-like.

Stoppa-type laparoscopic repair of complex groin defects.

With the laparoscopic approach, bilateral and complex groin defects can be corrected simultaneously by applying a preperitoneal mesh that covers the entire posterior wall of the groin, using a technique similar to the one described by Stoppa. We present our series of hernias repaired by the transabdominal preperitoneal laparoscopic approach with the Stoppa-type technique. The report consists of 78 cases of bilateral defects, of which 60% were indirect bilateral hernias, 23% direct bilateral, and 17% combined defects; 28.5% were recurrent hernias. Only minor complications were observed (hematomas and urinary retention) in some patients, but all resolved spontaneously. Three recurrences (0.7%) have been seen to this date. This method is recommended as the method of choice for complex groin defects.

Distribution of gonadotropin-releasing hormone immunoreactive systems in the brain of the Senegalese sole, Solea senegalensis.

The present paper reports the immunohistochemical distribution of the gonadotropin-releasing hormone (GnRH) structures in the brain of the Senegalese sole, Solea senegalensis. In this study, we have used two antibodies against the salmon GnRH and chicken GnRH-II forms and the streptavidin-biotin-peroxidase complex method. Immunoreactive cell bodies are observed at the junction between the olfactory bulbs and the telencephalon (terminal nerve ganglion cells), in the ventral telencephalon, in the preoptic parvocellular nucleus, and in the synencephalic nucleus of the medial longitudinal fasciculus. GnRH-immunoreactive fibres were found extensively throughout the brain, located in the telencephalon, preoptic area, hypothalamus, hypophysis, optic tectum, midbrain and rhombencephalon. The antisera used in this study against the two GnRH forms exhibited cross-reactivity on the same cell masses and did not allow cell populations expressing different GnRH forms to be discriminated clearly. However, anti-salmon GnRH immunostained the GnRH cells and fibres of the forebrain much more intensely, whereas the anti-chicken GnRH antiserum shows a higher immunoreactivity on synencephalic cells of the medial longitudinal fasciculus.