Towards a sustainable fisher-dolphin coexistence: Understanding depredation, assessing economic damage and evaluating management options.

Finding solutions for a sustainable coexistence between wildlife and humans is considered among the most challenging environmental management issues for scientists, conservationists, managers, and stockholders world-wide. Depredation by the common bottlenose dolphin (Tursiops truncatus) on small scale fisheries has increased in the recent years, leading to a growing conflict in many areas of the Mediterranean Sea and pressing for urgent management solutions. This study aims at developing a management framework for a sustainable coexistence between fishers and dolphins in Sardinia (Mediterranean Sea). Relying on the combination of different approaches (field study, literature review and Multi Criteria Decision Analysis), the scientific evidence necessary for understanding dolphin depredation were updated and improved, the related economic damage was calculated, and different management options were identified and evaluated by several stakeholder groups to support the decision-making process. Averaging for all investigated net types (gillnet and trammel net), a depredation frequency of 53% was found, the highest values ever found in both Sardinia and many other Mediterranean sites. Depredation probability was influenced by different factors, such as net type, fishing operation duration, depth of the fishing site and period. The estimated economic damage due to depredation ranges on average between 6492 and 11,925 euro per year and depends on the type of fishing net. The results from the field study, the literature review and the stakeholder involvement allowed us to define the most plausible and shared management options, identifying a framework for assessing and managing the conflict between fishers and dolphins for the creation of a more sustainable vision for the future.

Variations in coralligenous assemblages from local to biogeographic spatial scale.

The present study aims at contributing to the knowledge of the spatial variability of coralligenous reefs through the evaluation of patterns ranging from local to biogeographic scale around the island of Sardinia. The coralligenous reef assemblages of six areas were studied through a hierarchical sampling design: three sites per area were selected, in each site three plots were sampled and in each plot ten photographic samples were collected. The structure of coralligenous reefs across closed biogeographic regions is described, highlighting that nearly pristine assemblages, although characterized by similar high diversity, can be either dominated by animals, such as gorgonians and bryozoans, or macroalgae. The observed variations seem largely related to biogeographic patterns rather than spatial distance, supporting the need to identify specific reference conditions to assess the ecological quality of this habitat depending on the biogeographic area to be monitored.

Extreme events and conservation of subtidal habitats: Effects of a rainfall flood on coralligenous reefs.

The increase of the intensity and frequency of rainfall-dominated flood is considered a main effect of climate change. The present study evaluated the effect of a rainfall flood event on coralligenous reefs. The flooded site was compared to three control sites using a Before/After-Control/Impact (BACI) design. Sites were sampled using the STAR (STAndaRdized coralligenous evaluation procedure) approach and three ecological indices (ESCA, COARSE and ISLA) were calculated. At the disturbed site the number of species per sample, beta diversity, sensitivity levels of assemblages and the values of the three indices were lower after the flooding event, while the same variables did not decrease at the control sites. Algal turf and Dictyotales increased at the disturbed sites after the flood event, while Udoteaceae, erect sponges, bryozoans and Corallium rubrum decreased. This study provides evidence for identifying floods as a further cause of degradation for the coralligenous reef assemblages.

Macroalgal forest vs sea urchin barren: Patterns of macro-zoobenthic diversity in a large-scale Mediterranean study.

The study aimed at contributing to the knowledge of alternative stable states by evaluating the differences of mobile and sessile macro-zoobenthic assemblages between sea urchin barrens and macroalgal forests in coastal Mediterranean systems considering a large spatial scale. Six sites (100 s km apart) were selected: Croatia, Montenegro, Sicily (Italy), Sardinia (Italy), Tuscany (Italy), and Balearic Islands (Spain). A total of 531 taxa, 404 mobile and 127 sessile macro-invertebrates were recorded. Overall, 496 and 201 taxa were found in macroalgal forests and in barrens, respectively. The results of this large-scale descriptive study have met the expectation of lower macrofauna complexity and diversity in barrens rather than in macroalgal forests, and have allowed estimating the differences in levels of diversity and the consistency of variability across Mediterranean sites. Some peculiar patterns in barrens, related to both abundance of specific taxa and to high values of beta diversity, have been evidenced.

Benthic mucilage blooms threaten coralligenous reefs.

Mucilaginous aggregates produced by planktonic or benthic algae are considered ecological threats to marine systems. The study evaluated the effects of the spread of benthic mucilaginous aggregates on the structure of coralligenous assemblages. The assemblage and the quality of a site subjected to a benthic mucilage bloom were compared to those of two reference sites using a Before/After-Control/Impact (BACI) design. Results showed the α and ß-diversity, ESCA and COARSE quality ecological indices and the cover of encrusting algae and bryozoans were lower at the impact site after the mucilage event than at the control sites and at the impact site before the mucilage event. An opposite pattern was observed for the necrosis of gorgonians and the cover of algal turf. This study describes for the first time the impacts of ephemeral mucilage blooms on the whole coralligenous assemblage, identifying a further threat of this habitat and the need of adequate monitoring programs.

ALEX index enables detection of alien macroalgae invasions across habitats within a marine protected area.

A modified version of the ALien Biotic IndEX (ALEX) has been recently proposed to evaluate biological invasions in macroalgal assemblages. ALEX was applied in a Marine Protected Area where a recreational-fishing port is present testing the following hypotheses: ALEX increases with the distance from the port, it changes between the two directions off the port and it changes among three different habitats: Cystoseira beds, algal turf and dead matte of the seagrass Posidonia oceanica. A total of 78 native macroalgal taxa and 4 introduced species were found, the Chlorophyta Caulerpa cylindracea and the Rhodophyta Apoglossum gregarium, Acrothamnion preissii and Womersleyella setacea. All study sites were in high quality status highlighting that the assemblages investigated were at an early stage of NIS invasion. However, ALEX detected different values among conditions and habitats within the MPA, suggesting a local dynamics of NIS spread and different resistance to invasion of the investigated habitats.

The detrimental consequences for seagrass of ineffective marine park management related to boat anchoring.

Posidonia oceanica (L.) Delile meadows are recognized as priority habitat for conservation by the EU Habitats Directive. The La Maddalena Archipelago National Park (Mediterranean Sea) P. oceanica meadow, the dominant coastal habitat of the area, is mostly threatened by boat anchoring. 12 years after the establishment of mooring fields and anchoring restrictions, a study was conducted to measure their effectiveness on the conservation of seagrass and the mitigation of anchoring damage. We found that: (i) the condition of P. oceanica was disturbed, both in the mooring fields and in control locations; (ii) mooring fields and anchoring restrictions did not show to be an efficient system for the protection of seagrass, in fact anchor scars increased after the tourist season; (iii) the mooring systems had an impact on the surrounding area of the meadow, probably due to their misuse. On the basis of these results, management recommendations for marine parks are proposed.

Detection of low abundance mRNA of myeloid specific genes in cells of acute and chronic lymphoid leukemias by cRNA hybridization.

The hybridization to a complementary RNA (cRNA) probe both in situ and in solution was used to assay tiny amounts of mRNA of the lactoferrin (LF) and myeloperoxidase (MPO) genes in normal bone marrow cells and in acute and chronic lymphoid leukemias. Evidence is reported that this technique is much more sensitive than the standard Northern blot technique. The LF mRNA was detectable in three of seven cases of acute lymphoblastic leukemia (ALL) and in three of seven cases of chronic lymphocytic leukemia (CLL). Four cases of ALL were also positive when tested with the MPO cRNA. It is apparent from these results that myeloid specific mRNA, different from MPO, may be detected in leukemic cells with lymphoid phenotype using a method more sensitive than the Northern blot technique. Whether or not the molecular events observed in these cell populations reflect events physiologically occurring rather than a deregulation of gene expression associated to leukemogenesis remains to be established.

Ratios between the abundance of messenger RNA and the corresponding protein of two growth-related genes, c-myc and vimentin, in leukemia blast cells.

The abundance of the mRNAs of two growth-related genes, vimentin and c-myc, and that of the corresponding proteins have been studied in unstimulated and phytohemagglutinin-stimulated lymphocytes as well as in 18 populations of leukemic blast cells. The quantitative assay was carried out by densitometric scanning of Northern and Western blots. In normal lymphocytes the mRNA and the protein of both genes were almost undetectable. The phytohemagglutinin stimulation led to a sharp increase of the mRNA and the proteins of vimentin and c-myc. The increase was followed by a progressive fall of the gene products. The rate of decrease of the two mRNAs was similar to that of the corresponding proteins. In some leukemic populations very similar amounts of the vimentin protein were accompanied by amounts of the mRNA differing at least 25 times. Not unlikely, very similar amounts of p62c-myc corresponded to mRNA abundances differing at least 16 times. The coordinated biogenesis of both messenger RNAs and proteins, which occurs in mitogen-stimulated lymphocytes, is substituted, in approximately 30% of the leukemic blast cell populations, by molecular events leading to the accumulation of an excess of mRNA.

Overexpression of the MPO gene occurring in a case of APL without unusual genotypic characteristics.

Northern blot analysis of four typical cases of acute promyelocytic leukemia showed that one of the cell population examined was characterized by a very high level of expression of the myeloperoxidase (MPO) gene. Western blot analysis confirms that the protein content of the cells corresponded to the levels of the MPO mRNA. Southern blot studies of the DNA of this cell population ruled out the presence of any genome amplification or rearrangement. Chromosome hybridization studies in situ confirmed that the MPO gene was translocated on the long arm of chromosome 15. The observation that a typical genomic pattern may or may not be associated with the MPO overexpression leads us to believe that so far it is impossible to reach any conclusion about the significance of the translocation in the genesis of MPO overexpression.

[Expression of lactoferrin and myeloperoxidase genes in blasts of acute lymphoblastic leukemia]. / Espressione dei geni lattoferrina e mieloperossidasi in blasti di leucemie linfoblastiche acute.

The myeloperoxidase (MPO) and lactoferrin genes are strictly associated to the granulocytic differentiation pathway. Many reports show that immunologic and molecular cell markers, though to be lineage specific, are in fact expressed in cells of different lineages. We have identified lymphoid blast cell populations expressing the MPO gene at the mRNA level, but lacking any detectable MPO protein. We addressed this issue studying the expression of the MPO and lactoferrin genes in extremely homogeneous Acute Lymphoblastic Leukemia (ALL) blast cell populations. The study was carried out by solution hybridization of total cellular mRNA extracted from the leukemic populations with an antisense RNA transcribed by riboprobes. Either MPO or lactoferrin mRNAs were detectable in some ALL populations.

Expression of the myeloperoxidase gene in acute and chronic myeloid leukemias: relationship to the expression of cell cycle-related genes.

The expression of the myeloperoxidase (MPO) gene was studied, by means of Northern blot analysis in 14 cases of acute myeloid leukemia (AML), 11 cases of chronic myeloid leukemia (CML), and 6 cases of CML blast crisis, and in HL60 cells before and after induction of terminal differentiation with retinoic acid (RA), phorbol esters (TPA), or vitamin D. The expression of a panel of cell cycle-related genes, namely C-MYC, histone H3, ornithine decarboxylase, P53, vimentin, and calcyclin, was also studied in the same cell populations. Our results indicate that: (a) MPO gene expression (steady state mRNA levels) is strictly confined to the first stages of myeloid differentiation, reaching its peak at the promyelocyte stage and becoming undetectable in mature granulocytes and monocytes; (b) cells devoid of any detectable MPO enzymatic activity such as leukemic basophils have a high content of MPO mRNA; and (c) MPO gene expression is not related to the growth activity of the cell population. Finally, our results show that the pattern of expression of growth-regulated genes in the neoplastic myeloid disorders AML, CML, and CML blast crisis is remarkably different.

Effect of low frequency low energy pulsing electromagnetic field (PEMF) on X-ray-irradiated mice.

C3H/Km flora-defined mice were used to investigate the effect of exposure to pulsing electromagnetic field (PEMF) after total body x-ray irradiation. Prolonged exposure to PEMF had no effect on normal nonirradiated mice. When mice irradiated with different doses of x-ray (8.5 Gy, 6.8 Gy, and 6.3 Gy) were exposed to PEMF 24 h a day, we observed a more rapid decline in white blood cells (WBC) in the peripheral blood of mice exposed to PEMF at all the x-ray dosages used. No effect of exposure to PEMF was observed on the survival of the mice irradiated with 6.3 Gy and 8.5 Gy; in mice irradiated with 6.8 Gy, 2 out of 12 survived when exposed to PEMF as compared to 10 out of 12 control mice that were irradiated only. At day 4 after irradiation autoradiographic studies performed on bone marrow and spleen of 8.5-Gy-irradiated mice showed no difference between controls and mice exposed to PEMF, whereas on 6.8-Gy mice the bone marrow labeling index was lower in mice exposed to PEMF. In mice irradiated to 6.3 Gy we observed that the recovery of WBC in the peripheral blood was slowed in mice exposed to PEMF and their body weight was significantly lower than in control mice that were irradiated only. The spleen and bone marrow of the mice irradiated to 6.3 Gy and sacrificed at days 4, 14, 20, and 25 after irradiation were analyzed by autoradiography to evaluate the labeling index. Half of the spleens from mice sacrificed at day 25 after irradiation were used to evaluate the RNA content. Autoradiography showed that in the spleen and bone marrow of control mice, there were more cells labeled with [3H]thymidine at days 4 and 14 and less at days 20 and 25 after irradiation in comparison with mice irradiated and exposed to PEMF. The Northern blot analysis of histone H3 and p53 protein RNAs extracted from the spleens at day 25 after irradiation showed a slight increase in cycling cells among spleens of mice exposed to PEMF. We suggest that the exposure to PEMF immediately after x-ray irradiation results in increased damage.

[Polymerase chain reaction for the diagnostic identification of HIV infection]. / Reazione polimerasica a catena per la identificazione diagnostica di infezione da HIV.

The Acquired Immune deficiency Syndrome (AIDS), caused by the Human Immunodeficiency Virus (HIV) is now a worldwide social problem. Routine diagnostic procedures to identify infected individuals are based on the presence of antibodies against viral epitopes in the serum. There is nevertheless impelling need to detect directly the virus in people infected by HIV, independently of a serological response. In this study we describe the procedure which allows amplification of a specific segment of the HIV genome, through the Polymerase Chain Reaction (PCR), in infected individuals. This new approach represents a precious tool towards the diagnosis of HIV infection, it can be easily and quickly carried out on a large scale and will be capable of identifying HIV infected subjects before the development of antibodies.

Electrical stimulation of human femoral intertrochanteric osteotomies. Double-blind study.

Low-frequency pulsing electromagnetic fields (PEMF) are being used in nonunion healing at several centers around the world. Much debate exists about quantification of PEMF effects, especially in humans where no randomized studies have been performed. The results of a double-blind treatment of 32 consecutive patients treated with femoral intertrochanteric osteotomy for hip degenerative arthritis are reported. Roentgenographic evaluation and callus density measurements performed with an image analyzer showed a statistically significant difference between controls and stimulated patients (p less than 0.01). In this extremely homogeneous patient population, PEMF stimulation favored osteotomy healing.

Expression of oncogenes and cell cycle related genes in acute and chronic leukemias.

The authors have assayed the level of expression of several cell-cycle related genes in several populations of circulating myeloid leukemic blast cells. The genes explored included oncogenes such as c-myc, c-myb, p53, and cell-cycle-related genes such as vimentin, calcyclin, ornithine decarboxylase (ODC) and histone H3. Particular attention was given to analysis of the relationship existing between the mRNA levels of the histone H3 gene, which is expressed specifically in the S phase of the cell cycle, and the levels of other genes that are expressed in different stages of the G1 phase. Remarkable differences were observed among the different cases indicating that a differential expression of cell-cycle-related genes characterizes many acute leukemias. This differential expression is reflected in an altered ratio among G1-related genes and the H3 histone gene. The large fraction of leukemic cells which does not express histone H3 and therefore is functionally noncycling, shows a heterogeneous pattern of G1-related gene expression. This reflects the inability of most leukemic cells to progress through the G1 phase into the S phase of the cell cycle. This inability represents an abnormality of the cell cycle. It is concluded that the study of the expression of cell-cycle genes and protooncogenes in in understanding how leukemic cells enter a state of proliferation arrest, which appears to occur in a large fraction of leukemic cells.

Myeloperoxidase gene expression in blast cells with a lymphoid phenotype in cases of acute lymphoblastic leukemia.

By using a cDNA clone of the myeloperoxidase (MPO) gene, we have studied, by Northern blot analysis, the level of MPO mRNA in eight cases of acute lymphoblastic leukemia (ALL). The blast cell populations studied were characterized by morphologic, cytochemical, immunochemical, and molecular criteria. With all the methods used the populations were found to be highly homogeneous and showed a typical lymphoid phenotype. In particular, the Ig heavy-chain gene rearrangement was largely prevalent, and the germ line configuration was almost absent. However, in three of eight cases, high levels of MPO mRNA were detected. The remarkable homogeneity of the cell populations examined suggests that the MPO mRNA observed was present in cellular elements certainly identified as lymphoid. The absence of contamination by myeloid cells was confirmed by the results of Western blot analysis of the proteins of the cell population studied: no MPO protein was detectable. The levels of mRNA observed were high enough to be comparable to those observed in a promyelocytic cell population.

Slow healing fractures: can they be prevented? (Results of electrical stimulation in fibular osteotomies in rats and in diaphyseal fractures of the tibia in humans).

The purpose of the study was to evaluate the possibility of preventing delayed union in fractures by the use of low-frequency pulsing electromagnetic fields (PEMFs). The study was conducted in two parts, both with control groups. Fibular osteotomies in rats and diaphyseal fractures of the tibia in humans were treated with and without electrical stimulation (PEMF). The rats were sacrificed on the 8th and 23rd days respectively in order to evaluate the histological picture of the repair callus and its mechanical resistance. In the human subjects, the clinical and radiological follow-up took into account various factors known to affect the rate of union in the various fracture groups. The results obtained suggest that PEMF stimulation is capable of accelerating and modulating the physiological process of union by its favourable effect on osteogenesis.