New Tetrachromic VOF Stain (Type III-G.S) for Normal and Pathological Fish Tissues

10 páginas, 3 figuras, 1 tabla. Este trabajo esta dedicado a Sra. María Armenta.A new VOF Type III-G.S stain was applied to histological sections of different organs and tissues of healthy and pathological larvae, juvenile and adult fish species (Solea senegalensis; Sparus aurata; Diplodus sargo; Pagrus auriga; Argyrosomus regius and Halobatrachus didactylus). In comparison to the original Gutiérrez VOF stain, more acid dyes of contrasting colours and polychromatic/metachromatic properties were incorporated as essential constituents of the tetrachromic VOF stain. This facilitates the selective staining of different basic tissues and improves the morphological analysis of histochemical approaches of the cell components. The VOF Type III -6.5 stain is composed of a mixture of several dyes of varying size and molecular weight (Orange G<acid Fuchsin<Light green<Methyl Blue<Fast Green), which are used simultaneously, and it enables the individual tissues to be selectively differentiated and stained. Muscle fibers, collagen, reticulin and elastin fibers, erythrocytes, cartilage, bone, mucous cells, oocytes and larvae were selectively stained and differentiated. Dyes with small size and molecular weight (i.e Orange G), penetrate all tissue structures rapidly, but are only tightly retained in densely textured tissues (i.e erythrocytes). Methyl Blue is an interesting triarylmethane dye (large size and molecular weight), which is incorporated in this new VOF tetrachrome stain, and acquires histochemical significance when used at acid pH (2.8) because collagen and reticulin fibers, as well basophilic and metachromatic substances (strongly ionized sulphated glycoconjugates) can be identified. Muscle tissues show an evident green colour (Fast Green or Light Green affinities), even those isolated and/or diffuse muscle fibers present in the digestive submucosa layer. Connective tissues showed a specific and strong blue colour (Methyl Blue affinity) or mixed blue-red staining (Methyl Blue and Acid Fucshin affinities). Very noticeable is the staining of the mucous cells, as well as the hyaline capsule of the viral lymphocystic cells, which were stained blue-purple (carboxylated and/or strongly ionized sulphated groups). Cartilaginous tissues showed a blue or purple (Methyl Blue affinity) staining, and a specific red colour (Acid Fucshin affinity) was evident during calcification or in bone structures (i.e skeleton, fins, gills, teeth).This work (Spanish MCYT/AGL2003-03558).Peer reviewe

Course of infection with Lymphocystis disease virus in gilthead seabream (Sparus aurata)

Lymphocystis disease virus (LCDV) is the etiological agent of lymphocystis disease (LCD), a pathology that affects a wide variety of fish species. Data about LCDV pathogenesis are very short, and mainly limited to histopathological studies of skin lesions. Recent studies on viral genome detection (both by PCR or DNA-DNA in situ hybridization) suggest that LCDV establish a systemic and persistent infection in gilthead seabream (Sparus aurata), but further studies are necessary to prove if this infection is productive or not. In the present study viral quantification and viral mRNA detection (by qPCR and RT-qPCR) have been used to investigate LCDV multiplication in different organs of juvenile gilthead seabream. In addition, a histopathological study was carried out. Animals were collected from two commercial farms in Southwestern Spain. In one farm, where no LCD outbreaks have been recorded, apparently healthy fish were collected, whereas in the other farm, diseased and recovered (two months after LCD symptoms disappearance) fish were sampled. All the animals were LCDV-infected, and viral gene expression was detected in every organ analysed (caudal fin, intestine, liver, spleen, kidney and brain). In asymptomatic animals, both apparently healthy and recovered, a low-titre infection was observed, with the highest viral copy numbers detected in brain and kidney. In diseased fish, viral loads were significantly higher than in subclinical infected animals, being maximal in caudal fin, where lymphocysts were present in the dermis. Different histological alterations were observed in the internal organs from diseased fish analysed, although no hypertrophied cells were detected in any of them. In recovered fish, most of the organs examined presented similar histological features to those in healthy animals. Thus, pathological changes were only detected in the intestine and liver, although they were less severe than those observed in diseased fish. The results presented showed that LCDV establishes a systemic infection in juvenile gilthead seabream, which can be subclinical. In addition, although the disease is self-limiting, the virus is not removed after disease recovery, but produces a persistent infection.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tec

Manuel Gutiérrez-Rodríguez (1923-2009):A cytohistochemist from Cadiz

3 pages.-- Obituary.February 23, 2009 was the sad date on which Dr. Manuel Gutierrez Rodriguez passed away in his hometown of Cadiz, Spain. He was a scientist with a background of excellent, untiring research work; equally important, he was admired and warmly loved by his fellow colleagues as well as by many students at the Medical School and the Science School of the University of Cadiz, who over the years received and were influenced by his teaching.Peer reviewe

Manejo de la inestabilidad acromioclavicular crónica

ResumenLa articulación acromioclavicular representa el vínculo entre la clavícula y la escápula necesario para que se produzca una dinámica funcional sincronizada. La inestabilidad acromioclavicular crónica supone la producción de cambios en la orientación anatómica de la escápula, situación que condiciona alteraciones en la trayectoria cinemática de los grupos musculares que se pueden traducir finalmente en dolor crónico. Se han descrito una serie de estrategias quirúrgicas para el tratamiento de los pacientes con inestabilidad acromioclavicular crónica sintomática. El abanico de posibilidades contempla técnicas anatómicas y no anatómicas, cirugía abierta y cirugía asistida por artroscopia, injertos biológicos y plastias sintéticas. En el manejo quirúrgico de la inestabilidad acromioclavicular crónica de debe contemplar la reconstrucción de los ligamentos rotos, ya que se acepta que después de 3 semanas de producida la lesión dichas estructuras carecen de potencial de cicatrización.En el presente artículo se presenta una revisión de la literatura en cuanto al manejo de la inestabilidad acromioclavicular crónica.AbstractThe acromioclavicular joint represents the link between the clavicle and the scapula, responsible for the synchronized dynamics of the shoulder girdle. Chronic acromioclavicular joint instability involves the occurrence of changes in the anatomical orientation of the scapula, a situation that leads to changes in muscle kinematics that could result in chronic pain. Several surgical strategies for the management of patients with chronic and symptomatic acromioclavicular joint instability have been described. The range of possibilities includes anatomical and non-anatomical techniques, open and arthroscopy-assisted surgery, and biological and synthetic grafts. Surgical management of chronic acromioclavicular joint instability should involve the reconstruction of the torn ligaments, as it is accepted that after 3 weeks from the injury, these structures lack healing potential.This paper describes a review of the literature as regards to the management of chronic acromioclavicular instability

Management of chronic unstable acromioclavicular joint injuries

The acromioclavicular joint represents the link between the clavicle and the scapula, which is responsible for the synchronized dynamic of the shoulder girdle. Chronic acromioclavicular joint instability involves changes in the orientation of the scapula, which provokes cinematic alterations that might result in chronic pain. Several surgical strategies for the management of patients with chronic and symptomatic acromioclavicular joint instability have been described. The range of possibilities includes anatomical and non-anatomical techniques, open and arthroscopy-assisted procedures, and biological and synthetic grafts. Surgical management of chronic acromioclavicular joint instability should involve the reconstruction of the torn ligaments because it is accepted that from three weeks after the injury, these structures may lack healing potential. Here, we provide a review of the literature regarding the management of chronic acromioclavicular joint instability. Expert opinion, Level V

Histochemical characterisation of oocytes of the swordfish <i>Xiphias gladius</i>

This work was partially funded by ICMAN.CSIC and the Spanish projects (MCYT/AGL2005-02478 and AGL2006-13777- CO3-O2/ACU). J.B. Ortiz-Delgado is supported by the Programa Ramón y Cajal (MEC, Spain)

Reliability of preoperative measurement with standardized templating in Total Knee Arthroplasty

AIM: To investigate the correlation between preoperative measurement in total knee arthroplasty and the prosthetic size implanted. METHODS: A prospective double-blind study of 50 arthroplasties was performed. Firstly, the reliability and correspondence between the size of said measurement and the actual implant utilized was determined. Secondly, the existing correlation between the intra- and interobserver determinations with the intraclass correlation coefficient was analyzed. RESULTS: An overall correspondence of 54%, improving up to 92% when the measured size admitted a difference of one size, was found. Good intra- and interobserver reliability with an intraclass correlation coefficient greater than 0.90 (P < 0.001) was also discovered. CONCLUSION: Agreement between the preoperative measurement with standardized acetate templates and the prosthetic size implanted can be considered satisfactory. We thus conclude it is a reproducible technique

Design Basis Revision and Weak Link Analysis of LCV-115B/D

The objective of the present project is the evaluation of the actual operating conditions of a group of LCV-115B/D in case of design based accident, normal operating or testing. Within the analysis of the design conditions, the documentation must include pressure maximum differential expected during valve opening and closing, as well as pressure for both normal operations and abnormal events, as these operations are included in existing and approved design bases. Furthermore, weak link analysis of the critical components has been done to prove that such components can withstand new maximum design thrust requirements in these valves. In order to accomplish this objective, LCV-115B/D have been modeled using Spaceclaim code, and the stresses and strains within this valves have been analyzed using ANSYS code

Post-thaw quality assessment of testicular fragments as a source of spermatogonial cells for surrogate production in the flatfish Solea senegalensis

Cryopreservation of germ cells would facilitate the availability of cells at any time allow ing the selection of donors and maintaining quality control for further applications such as transplan tation and germline recovery. In the present study, we analyzed the efciency of four cryopreservation protocols applied either to isolated cell suspensions or to testes fragments from Senegalese sole. In tes tes fragments, the quality of cryopreserved germ cells was analyzed in vitro in terms of cell recovery, integrity and viability, DNA integrity (fragmentation and apoptosis), and lipid peroxidation (malondialde hyde levels). Transplantation of cryopreserved germ cells was performed to check the capacity of cells to in vivo incorporate into the gonadal primordium of Senegalese sole early larval stages (6 days after hatch ing (dah), pelagic live), during metamorphosis (10 dah) and at post-metamorphic stages (16 dah and 20 dah, benthonic life). Protocols incorporating dimethyl sulfoxide (DMSO) as a cryoprotectant showed higher number of recovered spermatogonia, especially in samples cryopreserved with L-15 + DMSO (0.39 ± 0.18 × 106 cells). Lipid peroxidation and DNA frag mentation were also signifcantly lower in this treat ment compared with other treatments. An important increase in oxidation (MDA levels) was detected in samples containing glycerol as a cryoprotectant, refected also in terms of DNA damage. Transplan tation of L-15 + DMSO cryopreserved germ cells into larvae during early metamorphosis (10 dah, 5.2 mm) showed higher incorporation of cells (27.30 ± 5.27%) than other larval stages (lower than 11%). Cryopreservation of germ cells using testes fragments frozen with L-15 + DMSO was demonstrated to be a useful technique to store Senegalese sole germline.LA/P/0101/2020info:eu-repo/semantics/publishedVersio

Cuantificación y alteraciones histopatológicas producidas por concentraciones subletales de cobre en Fundulus heteroclitus

25 páginas.[EN] Fed and starved Fundulus heteroclitus specimens were exposed for 30 days to sublethal copper concentrations (800 ?g Cu2+/L). The accumulation of copper and histopathological effects on the liver and gills were studied. In both fed and starved specimens, the liver showed a progressive copper accumulation during the experimental period (from 2 to 30 days), with higher levels of copper in the liver of starved fish. Copper accumulation was also detected in the gills, and the specimens exposed to copper for 15 to 30 days (both fed and starved) showed significant differences (P < 0.005) with respect to the control group and the 2-day treated fishes. The main histopathological alterations were: vacuolization of hepatic parenchyma, desquamation of gill epithelia, lamellar telangiectasia, as well as hepatic and branchial necrosis. The results of this paper provide evidence that the nutritional status of the fish has great importance in modifying its response to sublethal copper effects.[ES]Ejemplares de Fundulus heteroclitus alimentados y mantenidos en inanición fueron expuestos a concentraciones subletales de cobre (800 µg Cu2+/L) durante un periodo de 30 días. Se estudió la acumulación de cobre, así como las alteraciones histopatológicas producidas en las branquias y en el hígado. El hígado, tanto en los organismos alimentados como en los mantenidos en ayunas, presentó acumulación progresiva hasta los 30 días, con mayores niveles en ejemplares sometidos al ayuno. Por otro lado, en las branquias se detectó también acumulación de cobre, presentando el grupo de organismos expuestos de 15 a 30 días (alimentados y sometidos a inanición) diferencias significativas (P < 0.005) con respecto al grupo control y los tratados durante 2 días. Las principales alteraciones histopatológicas fueron: vacuolización del parénquima hepático, descamación del epitelio branquial, telangiectasia lamelar, así como necrosis branquial y hepática. Los resultados obtenidos en el presente trabajo muestran la importancia del estado nutricional de peces, como modificador de la respuesta de los mismos a los efectos subletales producidos por la exposición a cobre.Peer reviewe