Functional assessment of cryopreserved pig aortas for pharmaceutical research

Several in vitro studies have demonstrated diminished post-thaw functional activity. Therefore, the aim of this study was to investigate the consequences of thawing and storage method used on the post-thaw functional activity of cryopreserved pig aortas with the aim of adjusting the freezing and thawing protocol so that the vascular segments are preserved in the best possible state, maintaining structure and functionality so that they can later be transplanted with success. In vitro responses of frozen, thawed pig aortas were used to investigate the functional activity after thawing at 15 degrees C and 100 degrees C/min and after storage in gas or liquid phase of liquid nitrogen. Cryopreservation was performed in RPMI 1640 medium + 10% dimethylsulfoxide and the rate of cooling was -1 degrees C/min, until -150 degrees C was reached.After thawing the maximal contractile responses to all the contracting agonists tested (KCl, noradrenaline) were in the ranges of 13-27% compared with the responses in unfrozen pig aortas. Contractile responses were slightly better when thawing was performed at 15 degrees C/min compared with 100 degrees C/min. The endothelium independent relaxant responses to sodium nitroprusside were reduced ( P < 0.05). Cryostorage of pig arteries also resulted in a loss of the endothelium-dependent relaxant response to acetylcholine. The cryopreservation method used provided a limited preservation of pig aorta contractibility, a reduction of the endothelium independent relaxant responses, and no apparent preservation of the endothelium-dependent relaxation. It is possible that further refinements of the cryopreservation protocol might allow better post-thaw functional recovery of pig aortas.     

Functional assessment of cryopreserved human femoral arteries for pharmaceutical research

An established method for cryopreservation that might preserve the vascular and endothelial responses of human femoral arteries (HFAs) to be transplanted as allografts was studied. HFAs were harvested from multiorgan donors and stored at 4 degrees C in saline solution before cryostorage. Thirty HFA rings were isolated and randomly assigned to one control group of unfrozen HFAs (eight rings) and one group of cryopreserved HFAs (22 rings).Cryopreservation was performed in RPMI solution containing dimethylsulfoxide (DMSO) and the rate of cooling was -1 degrees C/min until -40 degrees C and faster rates until -150 degrees C was reached. The contractile and relaxant responses of unfrozen and frozen/thawed arteries were assessed in organ bath by measurement of isometric force generated by the HFAs.After thawing, the maximal contractile responses to the contracting agonist tested (noradrenaline) were in the range of 43% of the responses in unfrozen HFAs. The endothelium-independent responses to sodium nitroprusside were not altered whereas the endothelium-dependent relaxant responses to acetylcholine were weakly altered.The cryopreservation method used provided a limited preservation of contractility of HFAs, a good preservation of the endothelium-independent relaxant responses, and a good preservation of endothelium-dependent relaxation. It is possible that further refinements of the cryopreservation protocol, such as a slower rate of cooling and a more controlled stepwise addition of DMSO, might allow better post-thaw functional recovery.     

Functional assessment of cryopreserved human aortas for pharmaceutical research

We evaluated the impact of standard cryopreservation on functional properties of human aortic homografts. From seven human donors, the thoracic descending aorta was obtained. Effects of cryopreservation on contractibility and endothelium function were tested. After cryopreservation no endothelium-dependent or endothelium-independent relaxation was found and the contractibility was strongly affected. Arteries showed no function and loss of endothelial integrity after cryopreservation and thawing.     

Hypothalamic and hormonal control of the photoperiodically induced vernal functions in the White-crowned Sparrow,Zonotrichia leucophrys gambelii

To assess the role of the hypothalamo-hypophysial complex in the control of photoperiodically induced vernal premigratory responses in the White-crowned Sparrow, the effects of hypothalamic lesions and systemic administration of several hormones on these responses were investigated. Lesions that destroyed the posterior median eminence (PME) or the entire median eminence (ME) inhibited photoperiodically induced testicular growth, premigratory fattening and Zugunruhe. Lesions in the basal infundibular nucleus (IN) that resulted in complete inhibition of testicular growth abolished Zugunruhe, but allowed varying degrees of fattening. The systemic administration of prolactin, testosterone propionate (TP) or the combination thereof in the PME-lesioned birds induced fattening similar to that observed in photostimulated controls but did not induce Zugunruhe. It is concluded that testosterone and prolactin are the most important hormones involved in the control of vernal premigratory fattening. The role of these hormones in the induction of vernal Zugunruhe is not positively proven.     

Phase I/II study of immunotherapy with T-cell peptide epitopes in patients with stage IV melanoma

Previous studies in small groups of patients suggested that immunization of melanoma patients with peptide epitopes recognized by T cells could induce regression of melanoma. This approach was tested in 36 patients with stage IV melanoma. The (MHC class I–restricted) peptides were from gp100, MART-1, tyrosinase, and MAGE-3. The gp100 and MART-1 peptides had been modified to increase their immunogenicity. In half the patients (groups 3 and 4) the peptides were given in the adjuvant Montanide-ISA-720, and half the patients in both groups were given GM-CSF s.c. for 4 days following each injection. Treatment was well tolerated except for two severe erythematous responses to Montanide-ISA-720 and marked inflammatory responses at sites of GM-CSF administration in three patients. There were no objective clinical responses but stabilization of disease for periods from 3 to 12 months were seen in seven patients. Five of these were patients given the peptides in Montanide-ISA-720. Delayed-type hypersensitivity (DTH) skin test responses were also seen mainly in the patients given the peptides in Montanide-ISA-720. GM-CSF did not increase DTH responses in patients in the latter group but may have increased DTH responses in those not given peptides in Montanide-ISA-720. Inflammatory responses around s.c. metastases or regional lymph nodes were observed in two patients. These results suggest that the peptides are more effective when given in the adjuvant Montanide-ISA-720. Nevertheless, results from this study, together with those from a number of comparable studies, indicate that peptide vaccines are currently of minimal benefit to patients and support the need for ongoing development of new strategies in treatment of this disease.     

Control of development in plants and fungi by far-UV radiation

Far-UV (200–320 nm) radiation regulates development in plants and fungi. Some of these responses are controlled by a chromophore which absorbs strongly near 260 nm, possibly a nucleic acid. Other responses are controlled by a chromophore(s). with maximal in vivo sensitivity near 295 nm. In plants. far-UV induces genes in the phenylpropanoid pathway and the synthesis of phytoalexins and flavonoids. Far-UV also regulates growth rate. controls curvature and taxis, and stimulates sexual and asexual morphogenesis of fungi. Some of these developmental responses may prevent damage by far-UV radiation.     

Molecular communications between plant heat shock responses and disease resistance

As sessile, plants are continuously exposed to potential dangers including various abiotic stresses and pathogen attack. Although most studies focus on plant responses under an ideal condition to a specific stimulus, plants in nature must cope with a variety of stimuli at the same time. This indicates that it is critical for plants to fine-control distinct signaling pathways temporally and spatially for simultaneous and effective responses to various stresses. Global warming is currently a big issue threatening the future of humans. Reponses to high temperature affect many physiological processes in plants including growth and disease resistance, resulting in decrease of crop yield. Although plant heat stress and defense responses share important mediators such as calcium ions and heat shock proteins, it is thought that high temperature generally suppresses plant immunity. We therefore specifically discuss on interactions between plant heat and defense responses in this review hopefully for an integrated understanding of these responses in plants.     

Morphology,temperature and photoperiodic responses in Audouinella botryocarpa (Harvey) Woelkerling (Acrochaetiaceae,Rhodophyta) from Ireland

Abstract

Audouinella botryocarpa is reported for the first time from the British Isles. On the west and south coasts of Ireland and in the Isle of Man it grows in the intertidal on wave-splashed limpet shells and rock surfaces. Populations examined monthly in Galway Bay formed monosporangia throughout the year and tetrasporangia from December to March and May but no gametangial plants were found. Plants isolated into culture from monospores formed monosporangia at daylengths of 8, 10, 12, 14 and 16 h at 6.5, 8, 10, 11 and 15°C; the same plants formed tetrasporangia at daylengths ≤10 h at 8, 10 and 11°C but not at 6.5 and 15°C. Spores isolated from these plants gave rise to further monosporangial plants that also formed tetrasporangia under inductive conditions. Night-breaks of 1 h in a 16 h night prevented tetrasporangial reproduction and a critical daylength of ～10 h was found at 10°C. British Isles Audouinella botryocarpa typically has 6 rounded or irregularly-shaped chloroplasts per cell, each of which has a single, centrally-placed pyrenoid. The chloroplasts grow together in mature cells, typically giving the appearance of a single plate-like chloroplast with 6 pyrenoids. The implications of these observations for the generic classification of acrochaetioid algae is discussed and it is concluded that chloroplast morphology, number and the presence or absence of pyrenoids may provide the basis for a future generic realignment of acrochaetioid algae.     

The function of phytochrome A

Knowledge of the photoperceptive function of phytochrome A has improved substantially thanks to the availability of mutants lacking phytochrome A and transgenic plants transformed with the PHYA gene in sense or anti-sense orientation. In imbibed seeds, phytochrome A mediates very-low-fluence responses. In etiolated seedlings, phytochrome A mediates very-low-fluence responses, high-irradiance responses under continuous far-red light, responsivity amplification to phytochrome B and red-light enhancement of the phototropic response to blue light. In light-grown seedings, phytochrome A modulates the extent of response to reductions in red/far-red ratio perceived by phytochrome B, perceives daylength extensions and night interruptions affecting flowering, and perceives light treatments resetting endogenous rhythms. Under natural radiation these abilities are manifested during seed germination and seedling de-etiolation under dense canopies or extremely low light fluences, and during early neighbour detection, but other processes await experimental evaluation. Phytochrome A affects growth and development throughout the whole life cycle of angiosperms.     

Phase I/II study of treatment with dendritic cell vaccines in patients with disseminated melanoma

Previous studies have suggested that immunotherapy with dendritic cell (DC) vaccines may be effective in treatment of patients with AJCC stage IV melanoma. We examined this treatment in phase I/II studies in 33 patients with good performance status and low volume disease. Nineteen patients received DCs plus autologous lysates and 14 patients DCs plus peptides from the melanoma antigens MAGE-3.A2, tyrosinase, gp100, and MART-1. Keyhole limpet hemocyanin (KLH) was used as a helper protein and influenza peptide was given as a positive control. DCs were produced from adherent cells in blood lymphocytes (monocytic DCs), grown in IL-4 and GM-CSF without a maturation step. The DCs were injected into inguinal lymph nodes at weekly intervals (×4), 2 weeks (×1), and 4-weekly intervals (×2). There were 3 responses (3 partial responses) and 1 mixed response in the 19 patients treated with DCs plus autologous lysates. No responses were seen in the group treated with DCs plus peptides. Stable disease (defined as no progression over a period of 3 months) was seen in 4 patients in group 1 and 5 patients in group 2. Treatment was not associated with significant side effects. We examined whether DTH skin tests or assays of IFN- cytokine production may be useful predictors of clinical responses. Twenty-two of 30 patients had DTH responses to KLH and 12 of 13 patients had DTH responses to the influenza peptide. Five of 15 DTH responses were seen against autologous lysates. This was strongly correlated with clinical responses. Approximately half the patients had responses to MART-1 peptide and a third to the other melanoma peptides. Similarly, cytokine production assays showed responses to influenza in 7 of 13 patients, and approximately one third of patients had responses to the other peptides. No IFN- responses were seen in 5 patients against their autologous lysates. There was no correlation between assays of IFN- production and clinical responses. The present studies suggest that autologous lysates may be more effective than the melanoma peptides used in the study as the source of antigen for DC vaccines. DTH responses to autologous lysates appear useful predictors of clinical responses, but further work is needed to identify other measures associated with clinical responses.Abbreviations DC dendritic cells - DTH delayed hypersensitivity skin tests - KLH keyhole limpet hemocyanin - CTL cytotoxic T lymphocytes     

UV-B dose/dose-rate responses of seasonally abundant copepods of Puget Sound

Summary Refinements in modeling stratospheric processes and recent observational data have generated estimates of moderate future reductions in total global ozone. Nevertheless, considerable uncertainty and concern remain about whether or not the resulting increases in incident solar UV-B could increase the risk of harmful biological effects. The UV-B dose/dose-rate thresholds for five species of copepods collected at the surface in Puget Sound were determined and compared to previously studied zooplankton groups. Generally, species appearing later in the spring and summer were less UV-B sensitive. The UV-B daily-dose and dose-rate thresholds for the five species of copepods, while apparently less than present median incident levels, range from above to below estimated present and future subsurface (1 m) UV-B levels. A separate experiment with a relatively large copepod, Calanus pacificus, in which above and below threshold UV-B dose-rates were administered with various levels of photoreactivating light(UV-A and visible light), indicated the presence of photorepair and its full activation at relatively low intensity. To improve estimates of the extent to which present and predicted UV-B levels affect local populations of copepods and zooplankton, it is essential to more fully understant the natural conditions or processes which influence the composition and intensity of the UV irradiation these animals receive.Contribution No. 1760 from the School of Oceanography, University of Washington, Seattle, WA 98195, USA     

Osmotic responses of preimplantation mouse and bovine embryos and their cryobiological implications

Cells subjected to the events occurring before, during, and after freezing and thawing are exposed to major changes in the osmotic pressure of the surrounding medium; i.e., the osmolalities can exceed 30. An important question in understanding the mechanisms of injury is whether cells respond as ideal osmometers to these strongly anisotonic solutions. Mouse and bovine embryos from eight-cell to blastocyst stage were used to investigate the question. They were found to behave as ideal osmometers at room temperature over a wide range of tonicities; i.e., from four times isotonic to almost 1/3 times isotonic, ideality being defined by a Boyle-van't Hoff equation. Embryo volumes increased from 40 to 200% of isotonic over this range and survivals of mouse embryos were unaffected. However, outside this range the membrane apparently becomes leaky and the survival of mouse embryos drops sharply. Osmolalities rise to high values during freezing and the paper develops the thermodynamic equations to show how computed cell volumes as a function of subzero temperature can be translated into the Boyle-van't Hoff format of cell volume as a function of the reciprocal of osmolality.     

初级视皮层神经元响应反应与撤反应的朝向选择性

朝向选择性是初级视皮层(17区或V1)神经元的基本性质,在图形感知中起着关键作用.同时这些神经元对于持续时间大于100 ms的视觉刺激具有清晰的响应反应(Onset responses)和撤反应(Offset responses).以往的研究只关注响应反应的朝向选择性,而忽视了对撤反应的朝向选择性研究.我们比较了响应与撤反应的朝向调谐性质,大多数细胞的撤反应与响应反应基本上具有相似的最优朝向,而撤反应的朝向调谐宽度有窄于响应反应的趋势,撤反应的最优延迟普遍滞后于响应反应的最优延迟.撤反应的朝向选择性略强于响应反应和具有显著长的反应延迟提示,皮层内的反馈输入可能在形成撤反应的朝向选择性中起着作用.本研究揭示了撤反应的朝向选择性在刺激朝向的连续表征和主体在形状知觉的后期对朝向的精细区分中起着作用.     

Decreasing functional responses as a result of adaptive consumer behavior

Summary Several different mechanisms that may produce decreasing functional responses are investigated using models that assume that an optimally foraging consumer is exploiting one or two resources. Decreasing functional responses are associated with situations in which there are costs to resource consumption. If the process of resource acquisition has costs, decreasing functional responses may occur when there is a single homogeneous resource. If the cost is solely a function of the amount of resource ingested, decreasing functional responses on a single resource do not occur. Both types of cost can produce decreasing functional responses when there are two resource types and a trade-off relationship between consumption of one and consumption of the other. Decreasing functional responses seem to be most likely to occur on a food that yields high benefits and costs per unit of foraging time or effort when there is an alternative resource which yields low benefits and costs. Given this type of foraging choice, the functional response is most likely to decrease when the benefits of ingestion increase at a decreasing rate, and the costs of ingestion increase at an increasing rate with amount ingested. An important and unique consequence of decreasing functional responses is the possibility of population cycles in differential equation models of consumer-resource systems with non-reproducing resources; this is illustrated with a simple comsumer-resource model.     

Selection strategies and artificial evolution

Summary Artificial selection results in biolgical changes, creating artificial evolution. When using selection indexes, the artificial evolution depends on the relative economic (or other) weight of traits in the breeding objective, and on the phenotypic and genetic variances and covariances among these traits and the traits recorded in the selection index. As shown here, the selection strategy (in this case, individual selection versus progeny test selection) can also have marked effects on the kind of artificial evolution produced. Thus, where economic weights are uncertain, choice between alternative selection strategies might take into account the different types of animal or plant resulting.     

SEASONAL DIFFERENCES IN THE TEMPERATURE RANGES OF GROWTH OF VIRGINIA ALGAE1

One hundred and fifteen clonal, unialgal strains were isolated and tested for their ability to grow over a range of temperatures from 2 to 40° C. Responses of 63 strains isolated from habitats that were 6° C when sampled and 52 strains isolated from habitats that were 20° C when sampled showed trends toward increasing adaptation to cold or warm temperatures commensurate with their seasonal in situ temperatures. Based on temperature-growth responses alone, 24% of the plankton isolates and 17% of the periphyton isolates could be perennial within the natural habitats. At 5° C, 56% of the warm water plankton isolates and 48% of the warm water periphyton isolates were incapable of growth and, therefore, probably could not be important components of the winter algal community. Likewise at 25° C, 25% of the cold water plankton isolates and 13% of the cold water periphyton isolates were incapable of growth. Thus, temperature alone probably is an important variable regulating seasonal changes in algal community structure. Pollution of these habitats by a thermal enrichment averaging + 5° C year-round could effect a pronounced change in algal species composition because many more taxa could be perennial and more taxa would be incapable of growth during naturally warm periods.     

Consumer-food systems: why type I functional responses are exclusive to filter feeders

The functional response of a consumer is the relationship between its consumption rate and the abundance of its food. A functional response is said to be of type I if consumption rate increases linearly with food abundance up to a threshold level at which it remains constant. According to conventional wisdom, such type I responses are more frequent among filter feeders than among other consumers. However, the validity of this claim has never been tested. We review 814 functional responses from 235 studies, thereby showing that type I responses are not only exceptionally frequent among filter feeders but that they have only been reported from these consumers. These findings can be understood by considering the conditions that a consumer must fulfil in order to show a type I response. First, the handling condition: the consumer must have a negligibly small handling time (i.e. the time needed for capturing and eating a food item), or it must be able to search for and to capture food while handling other food. Second, the satiation condition: unless its gut is completely filled and gut passage time is minimal, the consumer must search for food at a maximal rate with maximal effort. It thus has to spend much time on foraging (i.e. searching for food and handling it). Our functional response review suggests that only filter feeders sometimes meet both of these conditions. This suggestion is reasonable because filter feeders typically fulfil the handling condition and can meet the satiation condition without losing time, for they are, by contrast to non-filter feeders, able simultaneously to perform foraging and non-foraging activities, such as migration or reproduction.     

Synergy of Interleukin (IL)-5 and IL-18 in eosinophil mediated pathogenesis of allergic diseases

Eosinophils are circulating granulocytes that have pleiotropic effects in response to inflammatory signals in the body. In response to allergens or pathogens, exposure eosinophils are recruited in various organs that execute pathological immune responses. IL-5 plays a key role in the differentiation, development, and survival of eosinophils. Eosinophils are involved in a variety of allergic diseases including asthma, dermatitis and various gastrointestinal disorders (EGID). IL-5 signal transduction involves JAK-STAT-p38MAPK-NFκB activation and executes extracellular matrix remodeling, EMT transition and immune responses in allergic diseases. IL-18 is a classical cytokine also involved in immune responses and has a critical role in inflammasome pathway. We recently identified the IL-18 role in the generation, transformation, and maturation of (CD101⁺CD274⁺) pathogenic eosinophils. In, addition, several other cytokines like IL-2, IL-4, IL-13, IL-21, and IL-33 also contribute in advancing eosinophils associated immune responses in innate and adaptive immunity. This review discusses with a major focus (1) Eosinophils and its constituents, (2) Role of IL-5 and IL-18 in eosinophils development, transformation, maturation, signal transduction of IL-5 and IL-18, (3) The role of eosinophils in allergic disorders and (4) The role of several other associated cytokines in promoting eosinophils mediated allergic diseases.     

Mitochondrial proteolysis: Its emerging roles in stress responses

Background

Mitochondria are multifunctional organelles that not only serve as cellular energy stores but are also actively involved in several cellular stress responses, including apoptosis. In addition, mitochondria themselves are also continuously challenged by stresses such as reactive oxygen species (ROS), an inevitable by-product of oxidative phosphorylation. To exert various functions against these stresses, mitochondria must be equipped with appropriate stress responses that monitor and maintain their quality.

Scope of review

Interestingly, increasing evidence indicates that mitochondrial proteolysis has important roles in mitochondrial and cellular stress responses. In this review, we summarize current advances in mitochondrial proteolysis-mediated stress responses.

Major conclusions

Mitochondrial proteases do not only function as surveillance systems of protein quality control by degrading unfolded proteins but also regulate mitochondrial stress responses by processing specific mitochondrial proteins.

General significance

Studies on the regulation of mitochondrial proteolysis-mediated stress responses will provide the novel mechanistic insights into the stress response research fields.     

Is the cardiac monitoring function related to the self in both the default network and right anterior insula?

The self has been proposed to be rooted in the neural monitoring of internal bodily signals and might thus involve interoceptive areas, notably the right anterior insula (rAI). However, studies on the self consistently showed the involvement of midline default network (DN) nodes, without referring to visceral monitoring. Here, we investigate this apparent discrepancy. We previously showed that neural responses to heartbeats in the DN encode two different self-dimensions, the agentive ‘I’ and the introspective ‘Me’, in a whole-brain analysis of magnetoencephalography (MEG) data. Here, we confirm and anatomically refine this result with intracranial recordings (intracranial electroencephalography, iEEG). In two patients, we show a parametric modulation of neural responses to heartbeats by the self-relatedness of thoughts, at the single trial level. A region-of-interest analysis of the insula reveals that MEG responses to heartbeats in the rAI encode the ‘I’ self-dimension. The effect in rAI was weaker than in the DN and was replicated in iEEG data in one patient out of two. We propose that a common mechanism, the neural monitoring of cardiac signals, underlies the self in both the DN and rAI. This might reconcile studies on the self highlighting the DN, with studies on interoception focusing on the insula.This article is part of the themed issue ‘Interoception beyond homeostasis: affect, cognition and mental health’.