Metabolite and transcriptome analysis of Campylobacter jejuni in vitro growth reveals a stationary-phase physiological switch.

Campylobacter jejuni is a prevalent cause of food-borne diarrhoeal illness in humans. Understanding of the physiological and metabolic capabilities of the organism is limited. We report a detailed analysis of the C. jejuni growth cycle in batch culture. Combined transcriptomic, phenotypic and metabolic analysis demonstrates a highly dynamic 'stationary phase', characterized by a peak in motility, numerous gene expression changes and substrate switching, despite transcript changes that indicate a metabolic downshift upon the onset of stationary phase. Video tracking of bacterial motility identifies peak activity during stationary phase. Amino acid analysis of culture supernatants shows a preferential order of amino acid utilization. Proton NMR (1H-NMR) highlights an acetate switch mechanism whereby bacteria change from acetate excretion to acetate uptake, most probably in response to depletion of other substrates. Acetate production requires pta (Cj0688) and ackA (Cj0689), although the acs homologue (Cj1537c) is not required. Insertion mutants in Cj0688 and Cj0689 maintain viability less well during the stationary and decline phases of the growth cycle than wild-type C. jejuni, suggesting that these genes, and the acetate pathway, are important for survival.

Behavioral and Physiological Responses to Light by the Sea Anemone Anthopleura elegantissima as Related to its Algal Endosymbionts

The endosymbiosis of algae with invertebrates may be viewed with at least two major orientations. On the one hand, one may focus on the plant and animal as essentially separate organisms living together, as the word symbiosis states. The products which are exchanged between the plant and animal and the effects of the association on either partner are then of particular interest. On the other hand, one may consider the partnership as an entity, and attempt to investigate the physiology, behavior, etc. of the symbiotic association, observing what differences may appear between the "plant-animal" and analogous non-symbiotic organisms. It is the second approach which I have tried to take in this thesis. I have concentrated on some effects of light on symbiotic and aposymbiotic sea anemones of the species Anthopleura elegantissima, particularly with respect to pigmentation and several types of behavior.

Role of G protein coupled inwardly rectifier K+ channels (GIRK) on the neurobiology depression

353 págs.

Environmentally induced physiological responses that determine fish survival and distribution: a review

Limitation to an aqueous habitat is the most fundamental physiological constraint imposed upon fish, phrases such as 'like a fish of water', convey our acceptance of the general unsuitability of fish for terrestrial existence. The constraints that restrict fish to an aquatic habitat relate to respiration, acid-base regulation, nitrogenous excretion, water balance and ionic regulation. A fish not adapted for an amphibious lifestyle when removed from water, becomes hypoxic and hypercapnic and soon succumbs to respiratory acidosis because the problem of excretion of H super(+) and C0 sub(2) are more immediate than lack of oxygen. This happen because fish gills collapse in air, while the ventilator arrangements that moves an incompressible medium (water) oven them become ineffective

MicroRNA-132 is a physiological regulator of hematopoietic stem cell function and B-cell development

MicroRNAs are a class of small non-coding RNAs that negatively regulate gene expression. Several microRNAs have been implicated in altering hematopoietic cell fate decisions. Importantly, deregulation of many microRNAs can lead to deleterious consequences in the hematopoietic system, including the onset of cancer, autoimmunity, or a failure to respond effectively to infection. As such, microRNAs fine-tune the balance between normal hematopoietic output and pathologic consequences. In this work, we explore the role of two microRNAs, miR-132 and miR-125b, in regulating hematopoietic stem cell (HSC) function and B cell development. In particular, we uncover the role of miR-132 in maintaining the appropriate balance between self-renewal, differentiation, and survival in aging HSCs by buffering the expression of a critical transcription factor, FOXO3. By maintain this balance, miR-132 may play a critical role in preventing aging-associated hematopoietic conditions such as autoimmune disease and cancer. We also find that miR-132 plays a critical role in B cell development by targeting a key transcription factor, Sox4, that is responsible for the differentiation of pro-B cells into pre-B cells. We find that miR-132 regulates B cell apoptosis, and by delivering miR-132 to mice that are predisposed to developing B cell cancers, we can inhibit the formation of these cancers and improve the survival of these mice. In addition to miR-132, we uncovered the role of another critical microRNA, miR-125b, that potentiates hematopoietic stem cell function. We found that enforced expression of miR-125b causes an aggressive myeloid leukemia by downregulation of its target Lin28a. Importantly, miR-125b also plays a critical role in inhibiting the formation of pro-B cells. Thus, we have discovered two microRNAs with important roles in regulating normal hematopoiesis, and whose dregulation can lead to deleterious consequences such as cancer in the aging hematopoietic system. Both miR-132 and miR-125b may therefore be targeted for therapeutics to inhibit age-related immune diseases associated with the loss of HSC function and cancer progression.

Sensitivity of sturgeons to environmental hypoxia: a review of physiological and ecological evidence

In this essay, three lines of evidence are developed that sturgeons in the Chesapeake Bay and elsewhere are unusually sensitive to hypoxic conditions: 1. In comparison to other fishes,sturgeons have a limited behavioral and physiological capacity to respond to hypoxia. Basal metabolism, growth, feeding rate, and survival are sensitive to changes in oxygen level, which may indicate a relatively poor ability of sturgeons to oxyregulate. 2. During summertime, temperatures >20°C amplify the effect of hypoxia on sturgeons and other fishes due to a temperature oxygen "squeeze" (Coutant 1987). In bottom waters, this interaction results in substantial reduction of habitat; in dry years, sturgeon nursery habitats in the Chesapeake Bay may be particularly reduced or even eliminated. 3. While evidence for population level effects due to hypoxia is circumstantial, there are corresponding trends between the absence of Atlantic sturgeon reproduction in estuaries like the Chesapeake Bay where summertime hypoxia predominates on a system-wide scale. Also, the recent and dramatic recovery of shortnose sturgeon in the Hudson River (4-bid increase in abundance from 1980 to1995) may have been stimulated by improvement of a large portion of the nursery habitat that was restored from hypoxia to normoxia during the period 1973-1978.

Investigations of certain eco-physiological characteristics of the broad and long-pincered crayfish in connection with their interspecific relationships. [Translation from: Lietuvas TSR Mokslu Akademijos Darbai, C Serija 2(40), 279-286, 1966.]

The results are described of eco-physiological investigations of the broad-pincered (Astacus astacus L.J.) crayfish and the long-pincered (Astacus astacus Esch.) crayfish, conducted in 1963—64 in the Institute of Zoology and Parasitology of the Academy of Science of the Lithuanian SSR, for the purpose of studying the interspecific relationship of these two species. In the course of the investigation were determined: the influence of the temperature of the environment on the consumption of oxygen by full grown individuals of both species and on the respiratory movements of the scaphognathites, the threshold temperatures and the saturation of the water by oxygen, the diel activity in the winter period.

Model systems for the study of drug hypersensitivity

I. It was not possible to produce anti-tetracycline antibody in laboratory animals by any of the methods tried. Tetracycline protein conjugates were prepared and characterized. It was shown that previous reports of the detection of anti-tetracycline antibody by in vitro-methods were in error. Tetracycline precipitates non-specifically with serum proteins. The anaphylactic reaction reported was the result of misinterpretation, since the observations were inconsistent with the known mechanism of anaphylaxis and the supposed antibody would not sensitize guinea pig skin. The hemagglutination reaction was not reproducible and was extremely sensitive to minute amounts of microbial contamination. Both free tetracyclines and the conjugates were found to be poor antigens.

II. Anti-aspiryl antibodies were produced in rabbits using 3 protein carriers. The method of inhibition of precipitation was used to determine the specificity of the antibody produced. ε-Aminocaproate was found to be the most effective inhibitor of the haptens tested, indicating that the combining hapten of the protein is ε-aspiryl-lysyl. Free aspirin and salicylates were poor inhibitors and did not combine with the antibody to a significant extent. The ortho group was found to participate in the binding to antibody. The average binding constants were measured.

Normal rabbit serum was acetylated by aspirin under in vitro conditions, which are similar to physiological conditions. The extent of acetylation was determined by immunochemical tests. The acetylated serum proteins were shown to be potent antigens in rabbits. It was also shown that aspiryl proteins were partially acetylated. The relation of these results to human aspirin intolerance is discussed.

III. Aspirin did not induce contact sensitivity in guinea pigs when they were immunized by techniques that induce sensitivity with other reactive compounds. The acetylation mechanism is not relevant to this type of hypersensitivity, since sensitivity is not produced by potent acetylating agents like acetyl chloride and acetic anhydride. Aspiryl chloride, a totally artificial system, is a good sensitizer. Its specificity was examined.

IV. Protein conjugates were prepared with p-aminosalicylic acid and various carriers using azo, carbodiimide and mixed anhydride coupling. These antigens were injected into rabbits and guinea pigs and no anti-hapten IgG or IgM response was obtained. Delayed hypersensitivity was produced in guinea pigs by immunization with the conjugates, and its specificity was determined. Guinea pigs were not sensitized by either injections or topical application of p-amino-salicylic acid or p-aminosalicylate.

Observations on the physiological development of trout eggs [Translation from: Roux. Arch. f. Entwicklungsmechanik der Organismen, 114. 771, 1929]

The osmotic pressure of the perivitelline fluid and the yolk of trout (Salmo trutta) eggs were measured separately by the Drucker-Schrein method. The permeability of the egg membrane and the variations in the osmotic pressure of the eggs when placed in salt solutions were also investigated.

Physiological aspects of the life cycle of the river lamprey, Lampetra fluviatilis L

The life cycle of the river lamprey, L. fluviatilis, is reviewed. The larval lamprey, or ammocoete, is a blind, filter-feeding animal, which normally lies concealed in the silt deposits of streams and rivers. After a period of 3-5 years in fresh water the ammocoete undergoes a metamorphosis in the summer months into a sexually immature, non-feeding stage known as the macrophthalia, which is active. This stage migrates downstream in late winter. It adopts a parasitic existence, in intertidal areas. After 18 months it returns to spawn in fresh water, after a final freshwater stage lasting up to 9 months. The river lamprey dies within a few days after the spawning period of 3-4 weeks, and none survive to spawn the following year.

Physiological ecology of the ciliated protozoon Loxodes

Loxodes faces special problems in living close to the oxic-anoxic boundary. In tightly-stratified ponds like Priest Pot its optimum environment may be quite narrow and it can be displaced by the slightest turbulence. Loxodes cannot sense an O sub(2) gradient directly but its ability to perceive gravity allows it to make relatively long vertical migrations. It is also sensitive to light and oxygen and it uses these environmental cues to modulate the parameters of its random motility: in the dark, it aggregates at a low O sub(2) tension and in bright light it aggregates in anoxic water. The oxic-anoxic boundary is also a zone where O sub(2) may be a scarce and transient resource, but Loxodes) can switch to nitrate respiration and exploit the pool of nitrate that often exists close to the base of the oxycline.

Apaf-1 Inhibitors Protect from Unwanted Cell Death in In Vivo Models of Kidney Ischemia and Chemotherapy Induced Ototoxicity

Background: Excessive apoptosis induces unwanted cell death and promotes pathological conditions. Drug discovery efforts aimed at decreasing apoptotic damage initially targeted the inhibition of effector caspases. Although such inhibitors were effective, safety problems led to slow pharmacological development. Therefore, apoptosis inhibition is still considered an unmet medical need. Methodology and Principal Findings: The interaction between Apaf-1 and the inhibitors was confirmed by NMR. Target specificity was evaluated in cellular models by siRNa based approaches. Cell recovery was confirmed by MTT, clonogenicity and flow cytometry assays. The efficiency of the compounds as antiapoptotic agents was tested in cellular and in vivo models of protection upon cisplatin induced ototoxicity in a zebrafish model and from hypoxia and reperfusion kidney damage in a rat model of hot ischemia. Conclusions: Apaf-1 inhibitors decreased Cytc release and apoptosome-mediated activation of procaspase-9 preventing cell and tissue damage in ex vivo experiments and in vivo animal models of apoptotic damage. Our results provide evidence that Apaf-1 pharmacological inhibition has therapeutic potential for the treatment of apoptosis-related diseases.