Thaw depth spatial and temporal variability at the Limnopolar Lake CALM-S site, Byers Peninsula, Livingston Island, Antarctica.

A new Circumpolar Active Layer Monitoring (CALM) site was established in 2009 at the Limnopolar Lake watershed in Byers Peninsula, Livingston Island, Antarctica, to provide a node in the western Antarctic Peninsula, one of the regions that recorded the highest air temperature increase in the planet during the last decades. The first detailed analysis of the temporal and spatial evolution of the thaw depth at the Limnopolar Lake CALM-S site is presented here, after eight years of monitoring. The average values range between 48 and 29cm, decreasing at a ratio of 16cm/decade. The annual thaw depth observations in the 100×100 m CALM grid are variable (Variability Index of 34 to 51%), although both the Variance Coefficient and the Climate Matrix Analysis Residual point to the internal consistency of the data. Those differences could be explained then by the terrain complexity and node-specific variability due to the ground properties. The interannual variability was about 60% during 2009-2012, increasing to 124% due to the presence of snow in 2013, 2015 and 2016. The snow has been proposed here as one of the most important factors controlling the spatial variability of ground thaw depth, since its values correlate with the snow thickness but also with the ground surface temperature and unconfined compression resistance, as measured in 2010. The topography explains the thaw depth spatial distribution pattern, being related to snowmelt water and its accumulation in low-elevation areas (downslope-flow). Patterned grounds and other surface features correlate well with high thaw depth patterns as well. The edaphic factor (E=0.05842m2/°C·day; R2=0.63) is in agreement with other permafrost environments, since frozen index (F>0.67) and MAAT (<-2°C) denote a continuous permafrost existence in the area. All these characteristics provided the basis for further comparative analyses between others nearby CALM sites.

Ganglio centinela en cáncer de mama: biopsia selectiva comparada con linfadenectomía axilar. Seguimiento a largo plazo / Sentinel ganglion in breast cancer: selective biopsy compared with axillary lymphadectomy

Fundamento. La biopsia selectiva del ganglio centinela(BSGC) ha reemplazado a la linfadenectomía axilar (LA),como procedimiento de elección, en el estadiaje del cáncerde mama en estadios iniciales y axila clínicamente negativa.El objetivo de este trabajo es comparar la supervivenciaglobal y libre de eventos de aquellas pacientes a lasque se realizó, en el periodo de validación de la técnica,BSGC seguida de LA respecto a las que se practicó BSGCy LA si el ganglio centinela (GC) presentaba metástasis.Material y métodos. Se han incluido 148 pacientes, 81pertenecientes al periodo de validación y 67 al grupo deaplicación clínica. El radiocoloide se administró intraperitumoralmente,obteniéndose imágenes hasta la visualizacióndel GC, posteriormente en la intervenciónquirúrgica se procedió a su identificación y extirpación.Resultados. En el grupo de validación, la eficacia de latécnica ha sido del 92,5%, la sensibilidad del 95,6% y latasa de falsos negativos del 4%. De las 81 pacientes, 75se encuentran libres de enfermedad (92,6%). De las 67pacientes pertenecientes al grupo de aplicación clínica,63 (94%) viven libres de enfermedad. Ninguna pacienteha presentado recurrencia ganglionar axilar.Conclusiones. En la validación de la técnica hemos obtenidounos valores que se hallan dentro de las exigenciasde calidad generalmente aceptadas. Con una mediade seguimiento de 6 años no hemos observado recurrenciaaxilar en ninguno de los dos grupos. No existediferencia estadísticamente significativa en la supervivenciaglobal y libre de eventos entre ambos grupos(AU)

Background. Selective biopsy of the sentinel ganglion(SBSG) has replaced axillary lymphadectomy (AL) asthe procedure of choice in staging breast cancer in itsinitial stages and in clinically negative axilla. The aimof this study is to compare global event-free survival ofthose patients subjected to SBSG followed by AL, duringthe period of validation of the technique, with respectto those subjected to SBSG and AL if the sentinelganglion (SG) showed metastasis.Methods. One hundred and forty-eight patients wereincluded, 81 belonging to the period of validation and67 to the clinical application group. Radiocoloid wasadministered intraperitumorally, obtaining images upuntil the visualisation of the SG; its identification andextirpation were carried out subsequently in the surgicalintervention.Results. The efficacy of the technique in the validationgroup was 92.5%, sensitivity was 95.6% and the rate offalse negatives was 4%. Of the 81 patients, 75 are freeof disease (92.6%). Of the 67 patients belonging to theclinical application group, 63 (94%) are free of disease.No patient has presented axillary ganglion recurrence.Conclusion. In the validation of the technique we obtainedvalues that fall within the demands of generallyaccepted quality. With an average follow up of 6 yearswe did not observe axillary ganglion recurrence in anyof the two groups. There is no statistically significantdifference in global and event free survival between thetwo groups(AU)

Effects of concentrated supernatants recovered from Lactobacillus plantarum on Escherichia coli growth and on the viability of a human promyelocytic cell line.

AIMS: The ability of concentrated supernatants from Lactobacillus plantarum to produce a disruption of plasma membrane in eukaryotic and prokaryotic cells has been examined. METHODS AND RESULTS: A strain of Lact. plantarum (tolerant to acid and bile salts and resistant to several antibiotics) was used. It inhibited the growth of pathogenic Escherichia coli and L. monocytogenes. Supernatants from Lact. plantarum were concentrated by centrifugation. Either E. coli or HL-60 cells (a human promyelocytic cell line) were treated in the presence of the concentrated supernatants. The effect of concentrated supernatants from Lact. plantarum on E. coli growth demonstrated a bacteriostatic activity and a loss of cell viability measured by sytox green staining. Concentrated supernatants were capable of disturbing plasma membrane in E. coli and of promoting a cytotoxic and lyctic action on HL-60 cells and on human erythrocytes, respectively. CONCLUSIONS: These results suggest that Lact. plantarum release an effective compound responsible for an important effect in the disruption of E. coli plasma membrane and for a cytototoxic activity on promyelocytic leukaemia cells. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first in vitro study about the antimicrobial and biological activities of concentrated supernatants from Lact. plantarum.

[Sentinel ganglion in breast cancer: selective biopsy compared with axillary lymphadectomy]. / Ganglio centinela en cáncer de mama: biopsia selectiva comparada con linfadenectomía axilar. Seguimiento a largo plazo.

BACKGROUND: Selective biopsy of the sentinel ganglion (SBSG) has replaced axillary lymphadectomy (AL) as the procedure of choice in staging breast cancer in its initial stages and in clinically negative axilla. The aim of this study is to compare global event-free survival of those patients subjected to SBSG followed by AL, during the period of validation of the technique, with respect to those subjected to SBSG and AL if the sentinel ganglion (SG) showed metastasis. METHODS: One hundred and forty-eight patients were included, 81 belonging to the period of validation and 67 to the clinical application group. Radiocoloid was administered intraperitumorally, obtaining images up until the visualisation of the SG; its identification and extirpation were carried out subsequently in the surgical intervention. RESULTS: The efficacy of the technique in the validation group was 92.5%, sensitivity was 95.6% and the rate of false negatives was 4%. Of the 81 patients, 75 are free of disease (92.6%). Of the 67 patients belonging to the clinical application group, 63 (94%) are free of disease. No patient has presented axillary ganglion recurrence. CONCLUSION: In the validation of the technique we obtained values that fall within the demands of generally accepted quality. With an average follow up of 6 years we did not observe axillary ganglion recurrence in any of the two groups. There is no statistically significant difference in global and event free survival between the two groups.

Analysis of caspase activities in rat mammary tumours induced by N-methyl-nitrosourea.

Normal breast development is controlled by a balance between cell proliferation and apoptosis. The balance between the two parameters is crucial for determining the growth or regression of breast tumours in response to therapies and treatments. Therefore, it is necessary to understand the role of apoptosis in tumour progression. Active caspases participate as essential elements in the execution of apoptotic mechanisms. In the present study, we analysed the activities of caspase-3, -8 and -9 as well as cytochrome c release in N-methyl-nitrosourea (NMU)-induced rat mammary tumours, in order to establish the apoptotic events that occur in tumour growth in this animal model. Forty female virgin Wistar rats were randomly divided into two groups. One group was injected intraperitoneally with three doses of 50 mg/kg body weight of NMU. The control group received the vehicle only. After 122 days of NMU injection, the rats were sacrificed and the tumours were excised and processed. Results showed that in mammary tumours induced by NMU, the apoptotic death receptor-mediated pathway is activated through caspase-3 and -8, but the apoptotic mitochondrial pathway is suppressed through a non-activating process of caspase-9 activity, despite the release of cytochrome c. In conclusion, these findings have demonstrated a suppression of the apoptotic mitochondrial pathway through a non-activating process of caspase-9 activity, despite the release of cytochrome c in mammary tumours induced by NMU. Although the apoptotic death receptor-mediated pathway is activated, it is not enough to maintain the balance between proliferation and apoptosis, and thus determine the overall growth of the tumour.

[Analysis of the immune resistance in an experimental murine model fed dietary lipids and infected with Listeria monocytogenes]. / Análisis de la resistencia inmune en un modelo murino experimental alimentado con dietas lipídicas e infectado con Listeria monocytogenes.

Several dietary lipids are capable of exerting an immunosupressor effect. This action may have undiserable effects on the host immune resistance to infectious diseases. The purpose of the present study was to determinate the immune status of mice fed dietary lipids and experimentally infected with a virulent strain of Listeria monocytogenes. Balb/c mice were divided into four groups and were fed with their respective diet: low fat diet (LF, 20%), olive oil diet (OO, 20%), fish oil diet (FO, 20%) and hydrogenated coconut oil (HCO, 20%). Mice were fed for four weeks and infected with L. monocytogenes by endovenous route. Results have shown a survival reduction in mice fed a diet containing FO, as well as a significant increase in the number of viable bacteria from spleen. In addition, we have observed an increase in the bactericidal activity in peritoneal cells from OO group, although the invasion of L. monocytogenes in cells from this group was larger. Finally, a significant reduction of lymphocyte proliferation was observed in the group fed an FO diet, whereas natural killer (NK) cell activity was not modified. These results indicate that dietary lipids constituted by polyunsaturated n-3 fatty acids reduce the murine immune resistance, whereas a diet constituted by OO-does not exert an immunosuppressor effect as relevant as FO diet, and it does not reduce the immune resistance leading to an efficient L. monocytogenes elimination.

Anti-oxidant properties of N-acetyl-L-cysteine do not improve the immune resistance of mice fed dietary lipids to Listeria monocytogenes infection.

BACKGROUND & AIMS: Current knowledge of the potential effects that several dietary lipids exert on immune functions indicates that these substances participate actively in the modulation of immune system by which they contribute to the improvement of the conditions of patients suffering from inflammatory disorders. However, long-chain n-3 polyunsaturated fatty acids induce an immunosuppressive status that leads to a reduction of the host natural resistance to infectious agents as well as to an enhancement of oxidative damage. Hence, the present study has been designed to evaluate the effects on the immune system of the antioxidant N-acetyl-L-cysteine (NAC) in mice fed dietary lipids and infected with Listeria monocytogenes. METHODS: Balb/c mice were fed for 4 weeks with diets containing either olive oil (OO, 20% by weight), fish oil (FO, 20% by weight) or hydrogenated coconut oil (HCO, 20% by weight). After dietary lipid administration mice were experimentally infected with L. monocytogenes or treated with NAC (25mg/ml intraperitoneally). RESULTS: NAC at a concentration of 1mM promoted a loss of cell viability, although no differences were observed among the four groups. After injection of NAC in combination with L. monocytogenes, 25% of mice fed a low-fat (LF) diet survived. However, in the groups fed dietary lipids no effect on survival of mice was found. NAC participated in the reduction of superoxide anion generation measured with nitroblue tetrazolium (NBT) in the group fed a FO diet. Finally, NAC reduced the recovery of L. monocytogenes from spleen of mice fed diets containing LF or HCO. CONCLUSIONS: On the basis of these results, we can confirm that the administration of NAC improves survival in mice fed LF diet, whereas a reduction in the generation of superoxide radicals was measured in mice fed a FO diet and infected with L. monocytogenes. Similarly, bacterial recovery was diminished in mice fed diets containing LF or HCO. Hence, these data reveal a beneficial effect of NAC in mice fed LF or HCO and a detrimental action of this antioxidant in mice fed diets containing FO or OO.

Modulatory effects of long-chain n-3 fatty acids on cell functions.

In recent years, it has been demonstrated that certain fatty acids are involved in the modulation of immune system functions. The mechanisms responsible for these effects are not fully elucidated, but many hypotheses have described numerous changes in the cell functionality as the main factors capable of altering the immune functions. In the present investigation, we have analysed the potential effects of FFA on cell viability, production of superoxide radicals or proteasome activity in assays in vitro. Thus, different FFA, such as OA, EPA or SA have been incorporated to cellular cultures at a concentration of 100 microM. Phospholipase, cyclooxygenase or lipooxygenase inhibitors abolished the loss of thymocyte viability exerted by EPA, the most immunosuppressive fatty acid. Similarly, measurement of the oxidative process by NBT reduction in cells treated with EPA was markedly increased. Nevertheless, the proteasome activity as a mechanism that participates in T-cell activation was not modified by direct action of the different fatty acids on the in vitro cultures. Overall, these results underline the differential role of several fatty acids (particularly long-chain n-3 polyunsaturated fatty acids) in order to modulate many functions of the immune system.

Immunomodulatory effects of dietary lipids alter host natural resistance of mice to Listeria monocytogenes infection.

Over the past two decades, unsaturated fatty acids have received particular attention due to their ability to suppress immune functions. Nevertheless, suppression of immune functions also involves a reduction of host natural resistance to eliminate the infectious agents. We have analyzed the role of dietary lipids on immune functions in cells cultured with Listeria monocytogenes. Bactericidal efficiency of peritoneal cells from mice fed a fish oil diet against this bacterium was reduced and the incubation of peritoneal cells with polyunsaturated fatty acids led to similar results. The levels of superoxide radicals in the presence of L. monocytogenes increased in cells from mice fed olive oil or fish oil diets. Proteasome activity, a mechanism that participates in T cell activation, was inhibited in all of the dietary groups assayed in the presence of L. monocytogenes, but this inhibition was abolished in the presence of both MG132 (a proteasome inhibitor) and L. monocytogenes. Overall, these results underline the potential role of fatty acids in the modulation of many functions of the immune system.

Loss of natural killer cell activity after murine tumor transplantation appears as a consequence of dietary lipid administration.

It is generally assumed that unsaturated fatty acids are important biological mediators that may be applied in the treatment of patients suffering from autoimmune disorders, in the modulation of tumorigenic processes or in the induction or inhibition of apoptosis. In this study, mice divided into three groups were fed dietary lipids; olive oil (20% w/w), fish oil (20% w/w) or hydrogenated coconut oil (20% w/w) for four weeks. Then, survival percentage, natural killer (NK) cell activity and lipid peroxidation were measured after murine lymphoma (LSTRA) transplantation. Survival percentage showed a reduced resistance to LSTRA tumor in mice fed fish oil diet and generaly loss of NK cell activity was observed after lymphoma transplantation. Lipid peroxidation from mice fed dietary lipids was measured in the presence of both phospholipase and cyclooxygenase inhibitors. In general, the data contained in this paper point to a detrimental effect caused by dietary lipids in mice transplanted with a lymphoma.

Potential intervention of Campylobacter jejuni in the modulation of murine immune response.

Campylobacter jejuni has been reported to produce different toxins that may modulate the immune response in both animals and humans. The effect of C. jejuni enterotoxin on the immune response was investigated in two groups of Balb/c mice. One of them was inoculated intraperitoneally with 1010 colony forming units (CFU) of an enterotoxigenic strain (CCUG 7580), and the second one with a non-enterotoxigenic strain (CCUG 7440). The number of polymorphonuclear (PMN) cells from spleen increased in both enterotoxigenic and non-enterotoxigenic strains as a consequence of C. jejuni infection. Notwithstanding, lymphocyte proliferation stimulated by lipopolysaccharide (LPS) was increased by both enterotoxigenic and non-enterotoxigenic strains. Interleukin-2 (IL-2) production from splenic cells was increased significantly by infection with the enterotoxigenic strain. Both enterotoxigenic and non-enterotoxigenic strains reduced the splenic response to sheep erythrocytes; the response was significantly suppressed for immunoglobulin M (Ig M) and for immunoglobulin G (Ig G) synthesis. These results suggest that C. jejuni is able to modify some components of the immune response in mice, and also that the enterotoxigenic strain has more immunomodulating activity than the non-enterotoxigenic strain.

Immune cell functions, lipids and host natural resistance.

Nutritional status may exert a profound effect on immune system functions. Hence, several parameters of immune system are modified by dietary lipid administration, as lymphocyte proliferation, cytokine production, natural killer activity, antigen presentation, etc. Thus, numerous studies have indicated the key role of lipids as immune response modulators. These properties have been applied in the treatment of autoimmune and inflammatory diseases. As a result, the reduction or suppression of immune status due to lipid incorporation promotes an impairment in the ability of host natural response to eliminate infectious microorganisms as bacteria or parasites. In the present review, we analyze the current status about the relationship among dietary lipids, reduction of immune parameters and reduction of host natural response against infectious diseases. Many discrepancies are discussed, although several studies indicate a close association between dietary lipid manipulation and impairment in the elimination of bacteria, viruses or parasites. On the other hand, other studies point out a beneficial effect of dietary lipid manipulation on the host natural response. Future investigations will determine the events involved in the regulation of immune response by fatty acids and their role in the elimination of pathogenic agents.

Modification of acquired immunity in BALB/c mice by aztreonam.

Recent studies have suggested that antibiotics may act as biological response modifiers. In this study we investigated the effect of aztreonam, a monobactam antibiotic, on different parameters of acquired immunity in BALB/c mice. Different dosages of aztreonam injected into mice induced an increase in the lymphoproliferative response to specific mitogens and in the production of interleukin-2 by splenic cells, as well as a decreased response of this immune population to sheep erythrocytes lower total blood cell counts and a lower percentage of monocytes than in untreated mice. These results show a modulatory action of aztreonam on different immune parameters, which is independent of its antimicrobial activity and that could be of interest in human therapy.

Enhanced resistance to experimental systemic candidiasis in tilorone-treated mice.

Candida albicans is an increasingly important opportunistic fungal pathogen in immunocompromised patients. Natural killer (NK) cells constitute an important immune effector mechanism and are involved in the response to different pathological disorders. We wished to determine if this immune mechanism is involved in the specific response to C. albicans. Tilorone hydrochloride and related compounds have been described to display antiviral and antitumoral activity, as well as to enhance NK cell activity. In this study, we show the antimicrobial activity of different tilorone analogues and the enhanced resistance of tilorone-treated mice in experimental systemic candidiasis. We also present data suggesting that there is a correlation between NK cell activation and the resistance to experimental systemic candidiasis. Thus, it seems that the immunosurveillance of metastatic spread and the infection by C. albicans share some immune effector mechanisms, in particular activation of NK cells.

Oxidation of a critical thiol residue of the adenine nucleotide translocator enforces Bcl-2-independent permeability transition pore opening and apoptosis.

Mitochondrial membrane permeabilization is a critical event in the process leading to physiological or chemotherapy-induced apoptosis. This permeabilization event is at least in part under the control of the permeability transition pore complex (PTPC), which interacts with oncoproteins from the Bcl-2 family as well as with tumor suppressor proteins from the Bax family, which inhibit or facilitate membrane permeabilization, respectively. Here we show that thiol crosslinking agents including diazenedicarboxylic acid bis 5N, N-dimethylamide (diamide), dithiodipyridine (DTDP), or bis-maleimido-hexane (BMH) can act on the adenine nucleotide translocator (ANT), one of the proteins within the PTPC. ANT alone reconstituted into artificial lipid bilayers suffices to confer a membrane permeabilization response to thiol crosslinking agents. Diamide, DTDP, and BMH but not tert-butylhydroperoxide or arsenite cause the oxidation of a critical cysteine residue (Cys 56) of ANT. Thiol modification within ANT is observed in intact cells, isolated mitochondria, and purified ANT. Recombinant Bcl-2 fails to prevent thiol modification of ANT. Concomitantly, a series of different thiol crosslinking agents (diamide, DTDP, and BMH, phenylarsine oxide) but not tert-butylhydroperoxide or arsenite induce mitochondrial membrane permeabilization and cell death irrespective of the expression level of Bcl-2. These data indicate that thiol crosslinkers cause a covalent modification of ANT which, beyond any control by Bcl-2, leads to mitochondrial membrane permeabilization and cell death.

Modulatory effects of dietary lipids on immune system functions.

Dietary lipid manipulation may affect a great number of immune parameters, such as lymphocyte proliferation, cytokine synthesis, natural killer (NK) cell activity, phagocytosis and so on. The immunomodulation induced by dietary fatty acids may be applied in the amelioration of inflammatory disorders, such as autoimmune diseases. However, the mechanisms that participate in these processes are still poorly understood. It is probable that modulation of immune system by fatty acids of the diet may occur by alteration of membrane fluidity, lipid peroxide formation, eicosanoid production or regulation of gene expression. However, recent studies have reported the effects of several free fatty acids on apoptosis induction of in vitro cultures. In fact, a possible explanation of the effects that fatty acids promote on the immune system cells could be associated with an apoptotic process performed in an irreversible way. In vivo studies have demonstrated the ability of fatty acids to alter the survival of animals fed diets containing oils and infected with a pathogenic bacterium. Experimental infection in animals fed dietary lipids produces a modification of resistance to micro-organisms. The present review analyses all of these parameters that dietary fatty acids are capable of altering in order to modify the immune response. Further studies will be needed to establish the mechanisms involved in immune system regulation, reduction of symptoms derived from autoimmune pathologies and so on.

Determination of natural resistance of mice fed dietary lipids to experimental infection induced by Listeria monocytogenes.

Current understanding based on the effect of dietary lipid manipulation upon immune system function indicates that fatty acids are involved in the modulation of the immune response through different and complex pathways. Reduction of several immune parameters by fatty acid action may be applied in the treatment of diseases characterised by an overactivation of the immune system. As a consequence, a reduction of host resistance against infectious agents has been reported in animals fed dietary lipids. The present study confirms the action of dietary lipids on the survival of mice infected with the pathogenic bacterium Listeria monocytogenes. A significant increase in peritoneal cells from mice fed a hydrogenated coconut oil diet was found, while a significant reduction of bacterial recovery from spleens of these mice was observed in this group. In addition, both eicosanoid and phospholipase inhibitors did not promote any modification of lymphocyte proliferation from mice fed olive oil or fish oil.

Modification of acquired immunity in mice by imipenem/cilastatin.

The immunomodulating properties of antimicrobial drugs may have important implications for clinical practice, particularly for those patients whose immune system has been compromised. In this study, we assessed the influence of different treatments with a beta-lactam antibiotic (imipenem/cilastatin) on several acquired immune responses of BALB/c mice; splenocyte responses to specific mitogens and to sheep red blood cells, IL-2 production and proportions of the different lympho-monocytic populations. Impenem/cilastatin was shown to modify some lymphocyte-associated immune functions and it would be useful to investigate whether immunomodulatory effects also occur in humans.

Effects of aztreonam on natural immunity in mice.

The influence of the dose and the duration of treatment with aztreonam, a monocyclic beta-lactam antibiotic, on the natural immune response of mice has been investigated. The results show the effects induced by the antibiotic on several immune parameters were affected by the duration of treatment. Thus, treatment with 28 mg/kg per day of aztreonam over 14 days increased every immune parameter tested, while treatment with 57 mg/kg per day of aztreonam for 7 days only enhanced the natural killer (NK) activity of splenocytes. Since aztreonam does not apparently impair the innate immune response, it might be a suitable therapy for the treatment of patients who are immunosuppressed.

Evaluation of cytokine production and phagocytic activity in mice infected with Campylobacter jejuni.

The effect of several Campylobacter jejuni strains on the immune response was analyzed in mice after intraperitoneal inoculation with 10(10) colony forming units (CFU). Three C. jejuni strains were assayed: CCUG 6968 (enterotoxigenic), CCUG 7580 (enterotoxigenic), and CCUG 7440 (non-enterotoxigenic). These C. jejuni strains induced a peritoneal inflammatory response and an important increase in the peritoneal phagocyte oxidative activity measured by chemiluminescence assay, as well as an increase in the number of peritoneal cells. Both interleukin-1 (IL-1) and tumor necrosis factor alpha (TNFalpha) production by peritoneal cells were not modified. However, C. jejuni 7440 caused a statistically significant increase in TNFalpha production. These results have demonstrated that different strains of C. jejuni induce an increase of the inflammatory response without a significant cytokine release. However, these infectious microorganisms may be eliminated efficiently by murine macrophages after phagocytosis.