RESUMO
BACKGROUND Molecular hydrogen (H2) has been widely reported to have benefiicial effects in diverse animal models and human disease through reduction of oxidative stress and inflammation. The aim of this study was to investigate whether hydrogen gas could ameliorate endotoxin-induced uveitis (EIU) in rats. MATERIAL AND METHODS Male Sprague-Dawley rats were divided into a normal group, a model group, a nitrogen-oxygen (N-O) group, and a hydrogen-oxygen (H-O) group. EIU was induced in rats of the latter 3 groups by injection of lipopolysaccharide (LPS). After that, rats in the N-O group inhaled a gas mixture of 67% N2 and 33% O2, while those in the H-O group inhaled a gas mixture of 67% H2 and 33% O2. All rats were graded according to the signs of uveitis after electroretinography (ERG) examination. Protein concentration in the aqueous humor (AqH) was measured. Furthermore, hematoxylin-eosin staining and immunostaining of anti-ionized calcium-binding adapter molecule 1 (Iba1) in the iris and ciliary body (ICB) were carried out. RESULTS No statistically significant differences existed in the graded score of uveitis and the b-wave peak time in the Dark-adapted 3.0 ERG among the model, N-O, and H-O groups (P>0.05), while rats of the H-O group showed a lower concentration of AqH protein than that of the model or N-O group (P<0.05). The number of the infiltrating cells in the ICB of rats from the H-O group was not significantly different from that of the model or N-O group (P>0.05), while the activation of microglia cells in the H-O group was somewhat reduced (P<0.05). CONCLUSIONS Post-treatment hydrogen gas inhalation did not ameliorate the clinical signs, or reduce the infiltrating cells of EIU. However, it inhibited the elevation of protein in the AqH and reduced the microglia activation.
Assuntos
Hidrogênio/uso terapêutico , Uveíte/terapia , Animais , Humor Aquoso/efeitos dos fármacos , Proteínas de Ligação ao Cálcio/efeitos dos fármacos , Corpo Ciliar/efeitos dos fármacos , Modelos Animais de Doenças , Endotoxinas/efeitos adversos , Hidrogênio/administração & dosagem , Hidrogênio/fisiologia , Iris/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Masculino , Proteínas dos Microfilamentos/efeitos dos fármacos , Microglia/efeitos dos fármacos , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Ratos , Ratos Sprague-Dawley , Uveíte/induzido quimicamenteRESUMO
Phosphorus deficiency in soil is one of the major limiting factors for plant growth. Plasma membrane H+-ATPase (PM H+-ATPase) plays an important role in the plant response to low-phosphorus stress (LP). However, few details are known regarding the action of PM H+-ATPase in mediating root proton (H+) flux and root growth under LP. In this study, we investigated the involvement and function of different Arabidopsis PM H+-ATPase genes in root H+ flux in response to LP. First, we examined the expressions of all Arabidopsis PM H+-ATPase gene family members (AHA1-AHA11) under LP. Expression of AHA2 and AHA7 in roots was enhanced under this condition. When the two genes were deficient in their respective Arabidopsis mutant plants, root growth and responses of the mutants to LP were highly inhibited compared with the wild-type plant. AHA2-deficient plants exhibited reduced primary root elongation and lower H+ efflux in the root elongation zone. AHA7-deficient plants exhibited reduced root hair density and lower H+ efflux in the root hair zone. The modulation of H+ efflux by AHA2 or AHA7 was affected by the action of 14-3-3 proteins and/or auxin regulatory pathways in the context of root growth and response to LP. Our results suggest that under LP conditions, AHA2 acts mainly to modulate primary root elongation by mediating H+ efflux in the root elongation zone, whereas AHA7 plays an important role in root hair formation by mediating H+ efflux in the root hair zone.
Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/fisiologia , Hidrogênio/fisiologia , Fósforo/deficiência , Raízes de Plantas/fisiologia , ATPases Translocadoras de Prótons/genética , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , ATPases Translocadoras de Prótons/metabolismo , Estresse FisiológicoRESUMO
Recent basic and clinical research has revealed that hydrogen is an important physiological regulatory factor with antioxidant, anti-inflammatory and anti-apoptotic protective effects on cells and organs. Therapeutic hydrogen has been applied by different delivery methods including straightforward inhalation, drinking hydrogen dissolved in water and injection with hydrogen-saturated saline. This review summarizes currently available data regarding the protective role of hydrogen, provides an outline of recent advances in research on the use of hydrogen as a therapeutic medical gas in diverse models of disease and discusses the feasibility of hydrogen as a therapeutic strategy. It is not an overstatement to say that hydrogen's impact on therapeutic and preventive medicine could be enormous in the future.
Assuntos
Pesquisa Biomédica/tendências , Gases/uso terapêutico , Hidrogênio/uso terapêutico , Animais , Anti-Inflamatórios/uso terapêutico , Antioxidantes/uso terapêutico , Indústria Química/métodos , Quimioprevenção/métodos , Modelos Animais de Doenças , Humanos , Hidrogênio/química , Hidrogênio/fisiologia , Proteínas Inibidoras de Apoptose/uso terapêutico , Modelos BiológicosRESUMO
PURPOSE OF REVIEW: This review presents our current understanding of the way metabolic acidosis induces calcium efflux from bone, and in the process, buffers additional systemic hydrogen ions associated with acidosis. RECENT FINDINGS: Acid-induced changes in bone mineral are consistent with a role for bone as a proton buffer. In response to metabolic acidosis in an in-vitro bone organ culture system, we observed a fall in mineral sodium, potassium, carbonate and phosphate, which each buffer protons and in vivo should increase systemic pH towards the physiologic normal. Initially, metabolic acidosis stimulates physicochemical mineral dissolution and subsequently cell-mediated bone resorption. Acidosis suppresses the activity of bone-resorbing cells, osteoblasts, decreasing gene expression of specific matrix proteins and alkaline phosphatase activity. There is concomitant acid stimulation of prostaglandin production by osteoblasts, which acting in a paracrine manner increases synthesis of the osteoblastic receptor activator of nuclear factor kappa B ligand (RANKL). The acid induction of RANKL then stimulates osteoclastic activity and recruitment of new osteoclasts to promote bone resorption and buffering of the proton load. Both the regulation of RANKL and acid-induced calcium efflux from bone are mediated by prostaglandins. SUMMARY: Metabolic acidosis, which occurs during renal failure, renal insufficiency or renal tubular acidosis, results in decreased systemic pH and is associated with an increase in urine calcium excretion. The apparent protective function of bone to help maintain systemic pH, which has a clear survival advantage for mammals, will come partly at the expense of its mineral stores.
Assuntos
Acidose/fisiopatologia , Reabsorção Óssea/fisiopatologia , Cálcio/fisiologia , Nefropatias/fisiopatologia , Acidose/etiologia , Densidade Óssea/fisiologia , Soluções Tampão , Proteínas de Transporte/fisiologia , Regulação da Expressão Gênica/fisiologia , Homeostase/fisiologia , Humanos , Hidrogênio/fisiologia , Concentração de Íons de Hidrogênio , Nefropatias/complicações , Glicoproteínas de Membrana/fisiologia , Osteoclastos/fisiologia , Prostaglandinas/fisiologia , Ligante RANK , Receptor Ativador de Fator Nuclear kappa-BRESUMO
Vitamin B6 is essential for cellular functions and growth due to its involvement in important metabolic reactions. Humans and other mammals cannot synthesize vitamin B6 and thus must obtain this micronutrient from exogenous sources via intestinal absorption. The intestine, therefore, plays a central role in maintaining and regulating normal vitamin B6 homeostasis. Due to the water-soluble nature of vitamin B6 and the demonstration that transport of other water-soluble vitamins in intestinal epithelial cells involves specialized carrier-mediated mechanisms, we hypothesized that transport of vitamin B6 in these cells is also carrier mediated in nature. To test this hypothesis, we examined pyridoxine transport in a model system for human enterocytes, the human-derived intestinal epithelial Caco-2 cells. The results showed pyridoxine uptake to be 1) linear with time for up to 10 min of incubation and to occur with minimal metabolic alteration in the transported substrate, 2) temperature and energy dependent but Na+ independent, 3) pH dependent with higher uptake at acidic compared with alkaline pHs, 4) saturable as a function of concentration (at buffer pH 5.5 but not 7.4) with an apparent Michaelis-Menten constant (Km) of 11.99 +/- 1.41 microM and a maximal velocity (Vmax) of 67.63 +/- 3.87 pmol. mg protein-1. 3 min-1, 5) inhibited by pyridoxine structural analogs (at buffer pH 5.5 but not 7.4) but not by unrelated compounds, and 6) inhibited in a competitive manner by amiloride with an apparent inhibitor constant (Ki) of 0.39 mM. We also examined the possible regulation of pyridoxine uptake by specific intracellular regulatory pathways. The results showed that whereas modulators of PKC, Ca+2/calmodulin (CaM), and nitric oxide (NO)-mediated pathways had no effect on pyridoxine uptake, modulators of PKA-mediated pathway were found to cause significant reduction in pyridoxine uptake. This reduction was mediated via a significant inhibition in the Vmax, but not the apparent Km, of the pyridoxine uptake process. These results demonstrate, for the first time, the involvement of a specialized carrier-mediated mechanism for pyridoxine uptake by intestinal epithelial cells. This system is pH dependent and amiloride sensitive and appears to be under the regulation of an intracellular PKA-mediated pathway.
Assuntos
Proteínas de Transporte/fisiologia , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Mucosa Intestinal/enzimologia , Piridoxina/metabolismo , Transdução de Sinais/fisiologia , Células CACO-2 , Relação Dose-Resposta a Droga , Humanos , Hidrogênio/fisiologia , Mucosa Intestinal/metabolismo , Transdução de Sinais/efeitos dos fármacos , Sódio/fisiologia , Trocadores de Sódio-Hidrogênio/fisiologiaRESUMO
To determine whether the excitabilities of pulmonary C fibers to chemical and mechanical stimuli are altered by CO(2)-induced acidosis, single-unit pulmonary C-fiber activity was recorded in anesthetized, open-chest rats. Transient alveolar hypercapnia (HPC) was induced by administering CO(2)-enriched gas mixture (15% CO(2), balance air) via the respirator inlet for 30 s, which rapidly lowered the arterial blood pH from a baseline of 7.40 +/- 0.01 to 7.17 +/- 0.02. Alveolar HPC markedly increased the responses of these C-fiber afferents to several chemical stimulants. For example, the C-fiber response to right atrial injection of the same dose of capsaicin (0.25-1.0 microg/kg) was significantly increased from 3.07 +/- 0.70 impulses/s at control to 8.48 +/- 1.52 impulses/s during HPC (n = 27; P < 0.05), and this enhanced response returned to control within approximately 10 min after termination of HPC. Similarly, alveolar HPC also induced significant increases in the C-fiber responses to right atrial injections of phenylbiguanide (4-8 microg/kg) and adenosine (0.2 mg/kg). In contrast, HPC did not change the response of pulmonary C fibers to lung inflation. Furthermore, the peak response of these C fibers to capsaicin during HPC was greatly attenuated when the HPC-induced acidosis was buffered by infusion of bicarbonate (1.36-1.82 mmol. kg(-1). min(-1) for 35 s). In conclusion, alveolar HPC augments the responses of these afferents to various chemical stimulants, and this potentiating effect of CO(2) is mediated through the action of hydrogen ions on the C-fiber sensory terminals.
Assuntos
Hidrogênio/fisiologia , Hipercapnia/fisiopatologia , Fibras Nervosas Amielínicas/efeitos dos fármacos , Fibras Nervosas Amielínicas/fisiologia , Alvéolos Pulmonares/fisiologia , Adenosina/farmacologia , Animais , Bicarbonatos/farmacologia , Biguanidas/farmacologia , Pressão Sanguínea/efeitos dos fármacos , Pressão Sanguínea/fisiologia , Capsaicina/farmacologia , Espaço Extracelular/metabolismo , Hidrogênio/metabolismo , Concentração de Íons de Hidrogênio , Pulmão/citologia , Pulmão/inervação , Masculino , Ratos , Ratos Sprague-Dawley , Mecânica Respiratória/efeitos dos fármacos , Mecânica Respiratória/fisiologia , Estimulação QuímicaRESUMO
The ion parametric resonance (IPR) model predicts that distinct patterns of field-induced biological responses will occur at particular magnetic field combinations which establish ion resonances. An important characteristic of resonance is the bandwidth response of the system, in part because it determines the required tolerances of the test system. Initial development of the IPR model used literature data to estimate the bandwidth for any ion resonance to be -/+10% of its exact resonance. Because the charge-to-mass ratio of hydrogen is much larger than any other biologically significant ion, hydrogen resonance provides a unique test case by which a single ionic bandwidth can be clearly measured. Of particular relevance is work by Trillo et al. that demonstrated a hydrogen-only, resonance-based IPR response of neurite outgrowth in PC-12 cells. The work reported here considers the response of nerve-growth-factor-stimulated PC-12 cells exposed to magnetic fields tuned at or near hydrogen resonance. This work was designed to test directly the IPR model hypothesis of a -/+10% ionic bandwidth. Consistent with the work of Trillo et al., resonance conditions were established using a 2.97 microT static magnetic field, and the AC frequency and field strength were varied to prove different aspects of the resonance. With this static field 45 Hz was the resonance frequency identified for hydrogen, 42.5 and 47.5 Hz were near-resonance frequencies, and 40 and 50 Hz bounded the assumed -/+10% hydrogen resonance bandwidth. We repeated each test three times. The cell responses at 45 Hz exposures agreed with the IPR model predictions and replicated results obtained by Trillo et al. Cells exposed to 42.5 and 47.5 Hz (near resonance) magnetic fields responded in the same general pattern as those exposed to 45 Hz fields, but neurite outgrowth was less than that observed at resonance. Measured results for cells exposed to either 40 Hz or 50 Hz fields were indistinguishable from off-resonance responses, consistent with the hypothesized bandwidth. These results confirm that the response bandwidth for the hydrogen ion is no greater than -/+10%, and give further support to the resonance-based predictions of the IPR model.
Assuntos
Hidrogênio/fisiologia , Magnetismo , Neuritos/fisiologia , Animais , Espectroscopia de Ressonância de Spin Eletrônica , Previsões , Modelos Biológicos , Fatores de Crescimento Neural/farmacologia , Neuritos/efeitos dos fármacos , Distribuição Normal , Células PC12 , Prótons , Ratos , Rotação , VibraçãoRESUMO
By using random mutagenesis and enrichment by chemostat culturing, we have developed mutants of Methanobacterium thermoautotrophicum that were unable to grow under hydrogen-deprived conditions. Physiological characterization showed that these mutants had poorer growth rates and growth yields than the wild-type strain. The mRNA levels of several key enzymes were lower than those in the wild-type strain. A fed-batch study showed that the expression levels were related to the hydrogen supply. In one mutant strain, expression of both methyl coenzyme M reductase isoenzyme I and coenzyme F420-dependent 5,10-methylenetetrahydromethanopterin dehydrogenase was impaired. The strain was also unable to form factor F390, lending support to the hypothesis that the factor functions in regulation of methanogenesis in response to changes in the availability of hydrogen.
Assuntos
Proteínas de Bactérias/metabolismo , Methanobacterium/isolamento & purificação , RNA Mensageiro/metabolismo , Monofosfato de Adenosina/análogos & derivados , Monofosfato de Adenosina/análise , Contagem de Colônia Microbiana , Flavinas , Hidrogênio/fisiologia , Methanobacterium/enzimologia , Methanobacterium/crescimento & desenvolvimento , Mutagênese , Riboflavina/análogos & derivados , Riboflavina/análiseRESUMO
The pathogenesis of pneumatosis cystoides coli remains obscure in the absence of an explanation for why pockets of gas should form in the first place and why they should be maintained in the wall and mesentery of the colon. Counterperfusion supersaturation could explain the formation and location of the gas cysts, which occur mostly near blood vessels on the mesenteric border of the colon, and the absence of methane gas in them. The hypothesis can be tested by treating patients with pneumatosis cystoides coli with heliox.
Assuntos
Doenças do Colo/etiologia , Hélio/uso terapêutico , Oxigênio/uso terapêutico , Pneumatose Cistoide Intestinal/etiologia , Pressão Atmosférica , Colo/irrigação sanguínea , Doenças do Colo/terapia , Difusão , Humanos , Hidrogênio/fisiologia , Modelos Biológicos , Nitrogênio/fisiologia , Pneumatose Cistoide Intestinal/terapiaRESUMO
Patients with pneumatosis cystoides intestinalis have been reported to excrete excessive H2 because of a lack of H2-consuming intestinal bacteria. This study describes a patient with bacterial overgrowth and pneumatosis of the small intestine whose colonic flora avidly consumed H2 but whose small bowel flora produced but did not consume H2. There is no commonly accepted mechanism whereby excessive luminal H2 causes intramural gas. An explanation is proposed in which an initial, transitory source of intramural gas is distinguished from the mechanism that results in the persistence of the gas. Independent of the initial source of gas, rapid diffusion of H2 from the lumen into an intramural gas bubble would cause N2, O2, and CO2 to diffuse from the blood into the bubble. As a result, the bubble would expand and then persist indefinitely as long as H2 continued to diffuse from the lumen to the intramural gas collection.
Assuntos
Hidrogênio/fisiologia , Pneumatose Cistoide Intestinal/metabolismo , Idoso , Bactérias/metabolismo , Testes Respiratórios , Colo/microbiologia , Humanos , Hidrogênio/metabolismo , Intestino Delgado/microbiologia , Masculino , Metano/metabolismo , Pneumatose Cistoide Intestinal/microbiologiaRESUMO
The phase resetting of the circadian oscillatory system by pulses of increased temperature (zeitgebers) and the temperature compensation of its period length during longer exposures are major features of the system, but are not well understood in molecular terms. In Neurospora crassa, the effects of pulses of increased temperature on the circadian rhythm of conidiation were determined and possible inputs to the oscillatory system tested, including changes in cyclic 3',5'-adenosine monophosphate (cAMP), inositol 1,4,5-trisphosphate and H+ concentrations, as well as changes of phosphorylation, synthesis and degradation of proteins. Following the kinetics of these parameters during exposure to increased temperature showed transient changes. Experimental manipulation of cAMP, Ca2+ and H+ levels, and of the synthesis and, possibly, degradation of proteins, resulted in phase shifts of the oscillatory system. It is assumed that the temperature signal affects the oscillator(s) by multiple pathways and shifts the whole state of the oscillatory system. Second messenger levels, protein synthesis and protein degradation show adaptation to longer exposures to elevated temperature which may be involved in the temperature compensation of the period length. The temperature compensation is also proposed to involve a shift in the state of all or most oscillator variables.
Assuntos
Ritmo Circadiano/fisiologia , Neurospora crassa/fisiologia , Cálcio/fisiologia , AMP Cíclico/fisiologia , Hidrogênio/fisiologia , Transporte de Íons , TemperaturaRESUMO
H and K ions play central roles in prorenin processing and secretion, and prorenin is abnormally expressed in H and K disorders. At the surface membrane of juxtaglomerular (JG) cells, K is sensed and regulated by K channels (coupled to Cl channels and activated by excess Ca), Na-K-adenosinetriphosphatase, and a KCl/H exchange transporter (regulated by Ca). In JG cell granular membrane, K flux is regulated by K channels and a KCl/H exchange transporter (activated by Ca). H channels and a H pump reside in the granular membrane, which maintain H concentration in the granular matrix at least two orders of magnitude greater than in cytosol. The H pump may also be responsible for maintaining the acidic matrix required for maximal prorenin processing to renin by prohormone convertase for human renin (PCren), the prorenin convertase. These molecules form the core of a chemiosmotic system, which appears to regulate both prorenin processing and renin secretion. Renin secretion and prorenin processing appear to be of more than causal significance in clinical disorders characterized by chemiosmotic imbalance. A critical review of the literature supports the following general conclusions. First, hyperrenin state defines the initial phase in the pathogenesis of heart disease, diabetes mellitus, and hypertension. Second, low-renin syndrome defines the transition-to-establish phase in the pathogenesis of heart disease, diabetes mellitus, and hypertension in which the key feature is renin secretory hyporesponsivity. Third, renin disorders are usually associated with other endocrine disorders (polyendocrinopathies types I, II, and III), suggesting that renin may be an important molecule in the processing of chemiosmotic forces. The key chemiosmotic molecules (K and H) are also important in the processing and export of most (if not all) hormones. Thus, by regulating K and H homeostasis, renin may regulate the endocrine system.
Assuntos
Precursores Enzimáticos/metabolismo , Hidrogênio/fisiologia , Potássio/fisiologia , Processamento de Proteína Pós-Traducional , Renina/metabolismo , Animais , Doenças do Sistema Endócrino/metabolismo , HumanosRESUMO
The Ca(2+)-extruding ATPase pump of the human platelet was studied in situ by measuring Ca2+ extrusion from quin2-overloaded platelets (Johansson, J.S., Haynes, D.H. 1988. J. Membrane Biol. 104:147-163). Cytoplasmic pH (pHcyt) was measured by BCECF fluorescence in parallel experiments. The pump was studied by raising the cytoplasmic free Ca2+ to 2.5 microM and monitoring active Ca2+ extrusion into a Ca(2+)-free medium. The pump was shown to perturb pHcyt, to not respond to changes in membrane potential and to respond to imposed changes in pHcyt in a manner consistent with the Ca2+ pump acting as a 2 Ca2+/nH+ exchanger. (i) Raising the external pH (pHext) from 7.40 to 7.60 lowers the Vmax of the pump in basal condition (Vmax,1) from 110 +/- 18 to 73 +/- 12 microM/min (= mumol/liter cell volume/min). (ii) Lowering pHext to 7.13 raised Vmax,1 to 150 +/- 15 microM/min. (iii) In an N-methyl-D-glucamine (NMDG+) medium, the pump operation against high [Ca2+]cyt acidifies the cytoplasm by -0.36 +/- 0.10 pH units, and the pump becomes self-inhibited. (iv) Use of nigericin to drive pHcyt down to 6.23 reduces the Vmax,1 to 18 +/- 11 microM/min. (v) Alkalinization of the cytoplasm by monensin in the presence of Na+ raises the Vmax,1 (basal state with Km,1 = 80 nM) to 136 +/- 24 microM/min, but also activates the pump fourfold (Vmax,2 = 280 +/- 28 microM/min; Km,2 = 502 +/- 36 nM). (vi) Transient elevation of pHcyt by NH4Cl at high [Ca2+]cyt activates the pump eightfold (Vmax,2 > or = 671 +/- 350 microM/min). The large activation by alkaline pHcyt at high [Ca2+]cyt can be explained by Ca(2+)-calmodulin activation of the pump (Valant, P.A., Adjei, P.N., Haynes, D.H. 1992. J. Membrane Biol. 130:63-82) and by increased Ca2+ affinity of calmodulin at high pH.
Assuntos
Plaquetas/enzimologia , Canais de Cálcio/fisiologia , ATPases Transportadoras de Cálcio/fisiologia , Citoplasma/fisiologia , Plaquetas/fisiologia , Cálcio/sangue , Cálcio/fisiologia , Canais de Cálcio/sangue , ATPases Transportadoras de Cálcio/sangue , Citoplasma/metabolismo , Humanos , Hidrogênio/metabolismo , Hidrogênio/fisiologia , Concentração de Íons de Hidrogênio , CinéticaRESUMO
In Syrian hamster embryo cells, intracellular acidification (but not alkalization) results in proliferation, immediate-early-gene expression and tyrosine phosphorylation. In addition, both intracellular acidification and alkalization result in serine/threonine phosphorylation and de novo protein synthesis of specific proteins. Calcium is not mobilized in response to either intracellular alkalization or acidification. Neither intracellular acidification nor alkalization altered the serum proliferative signal while intracellular alkalization (but not acidification) reduced the epidermal-growth-factor-induced proliferative signal, tyrosine phosphorylation and immediate-early-gene expression. Finally, intracellular acidification (but not alkalization) could induce immediate-early-gene expression in cells growing in the presence of serum, indicating that the pH signalling pathway is not down modulated by the serum signalling pathway. These results, while indirect, indicate that hydrogen ions may play an important role in mitogen-signal transduction in Syrian hamster embryo cells.
Assuntos
Hidrogênio/fisiologia , Proteínas/metabolismo , Transdução de Sinais/fisiologia , Animais , Cálcio/metabolismo , Divisão Celular , Células Cultivadas , Cricetinae , Fator de Crescimento Epidérmico/farmacologia , Feminino , Expressão Gênica , Genes fos , Genes myc , Concentração de Íons de Hidrogênio , Íons , Fosforilação , Gravidez , Biossíntese de Proteínas , Proteínas/genéticaRESUMO
Bradyrhizobium japonicum expresses hydrogenase in microaerophilic free-living conditions in the presence of nickel. Plasmid-borne hup-lacZ transcriptional fusion constructs were used to study the regulation of the hydrogenase gene. The hydrogenase gene was transcriptionally induced under microaerobic conditions (0.1 to 3.0% partial pressure O2). The hydrogenase gene was not transcribed or was poorly transcribed in strictly anaerobic conditions or conditions above 3.0% O2. Hydrogen gas at levels as low as 0.1% partial pressure induced hydrogenase transcription, and a high level of transcription was maintained up to at least 10% H2 concentration. No transcription was observed in the absence of H2. Hydrogenase was regulated by H2, O2, and Ni when the 5'-upstream sequence was pared down to include base number -168. However, when the upstream sequence was pared down to base number -118, the regulatory response to O2, H2, and Ni levels was negated. Thus, a common cis-acting regulatory region localized within 50 bp is critical for the regulation of hydrogenase by hydrogen, oxygen, and nickel. As a control, the B. japonicum hemA gene which codes for delta-aminolevulinic acid synthase was also fused to the promoterless lacZ gene, and its regulation was tested in the presence of various concentrations of O2 and H2. hemA-lacZ transcription was not dependent on levels of Ni, O2, or H2. Two different hup-lacZ fusions were tested in a Hup- background, strain JH47; these hup-lacZ constructs in JH47 demonstrated dependency on nickel, O2, and H2, indicating that the hydrogenase protein itself is not a sensor for regulation by O2, H2, or nickel.
Assuntos
Hidrogênio/fisiologia , Hidrogenase/genética , Níquel/fisiologia , Oxigênio/fisiologia , Regiões Promotoras Genéticas , Rhizobiaceae/genética , Transcrição Gênica , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Óperon Lac , Regiões Promotoras Genéticas/efeitos dos fármacos , Proteínas Recombinantes de Fusão/genética , Rhizobiaceae/efeitos dos fármacos , Rhizobiaceae/enzimologia , Transcrição Gênica/efeitos dos fármacosAssuntos
Acidose Tubular Renal/fisiopatologia , Adenosina Trifosfatases/fisiologia , Anidrases Carbônicas/fisiologia , Rim/fisiopatologia , ATPase Trocadora de Sódio-Potássio/fisiologia , Equilíbrio Ácido-Base/fisiologia , Animais , ATPase Trocadora de Hidrogênio-Potássio , Humanos , Hidrogênio/fisiologia , Capacidade de Concentração Renal/fisiologia , Néfrons/enzimologiaRESUMO
The plasma membranes of most if not all vertebrate cells contain a transport system that mediates the transmembrane exchange of sodium for hydrogen. The kinetic properties of this transport system include a 1:1 stoichiometry, affinity for lithium and ammonium ion in addition to sodium and hydrogen, the ability to function in multiple 1:1 exchange modes involving these four cations, sensitivity to inhibition by amiloride and its analogues, and allosteric regulation by intracellular protons. The plasma membrane sodium-hydrogen exchanger plays a physiological role in the regulation of intracellular pH, the control of cell growth and proliferation, stimulus-response coupling in white cells and platelets, the metabolic response to hormones such as insulin and glucocorticoids, the regulation of cell volume, and the transepithelial absorption and secretion of sodium, hydrogen, bicarbonate and chloride ions, and organic anions. Preliminary evidence raises the possibility that the sodium-hydrogen exchanger may play a pathophysiological role in such diverse conditions as renal acid-base disorders, essential hypertension, cancer, and tissue or organ hypertrophy. Thus, future research on cellular acid-base homeostasis in general, and on plasma membrane sodium-hydrogen exchange in particular, will enhance our understanding of a great variety of physiological and pathophysiological processes.
Assuntos
Proteínas de Transporte/fisiologia , Membrana Celular/fisiologia , Hidrogênio/fisiologia , Sódio/fisiologia , Equilíbrio Ácido-Base , Animais , Transporte Biológico Ativo , Plaquetas/fisiologia , Cálcio/fisiologia , Proteínas de Transporte/antagonistas & inibidores , Cátions Monovalentes/metabolismo , Epitélio/metabolismo , Hormônios/fisiologia , Humanos , Concentração de Íons de Hidrogênio , Hipertensão/fisiopatologia , Leucócitos/fisiologia , Proteínas de Membrana/metabolismo , Neoplasias/fisiopatologia , Trocadores de Sódio-Hidrogênio , ATPase Trocadora de Sódio-Potássio/metabolismo , Equilíbrio HidroeletrolíticoRESUMO
Phospholipase C (EC 3.1.4.3.) from Clostridium perfringens (crude extracts) was used to study the role of phospholipids in the osmotic permeability of the urinary bladder of the toad. When added to the serosal bath (430 mU/ml) it inhibited the effects of antidiurectic hormone (ADH) and exogenous cyclic AMP. Under the same conditions the increase in osmotic flow produced by serosal hypertonicity (SH) was slightly enhanced by the lipase. The hydroosmotic effect of SH was greatly potentiated by the lipase by decreasing 10-fold the Ca2+ concentration. The SH-induced flow was inhibited by the lipase if the Ca2+ or the H+ concentration was increased 10-fold, but not if the increase in positive charges was produced by a concentration of Mg2+. Phospholipase C had no effect on the action of either ADH or SH if added to the mucosal bath. Serosal neuraminidase or phospholipase A2 could not mimic the effect of phospholipase C on SH. The effect of phospholipase C on the response to SH was not modified if fatty acid-free bovine serum albumin was added to the bath. Therefore, the release of products of lipolysis into the bath do not seem to be responsible for the effects of phospholipase C on SH-induced water flow. The results suggest that the effects of the enzyme on the composition and rearrangement of lipids at the basolateral membrane produce modifications of the water flow. Ca2+ and H+ may modify the enzyme-substrate interaction, suggesting that different phospholipids may be differentially involved in the control of water permeability of the basolateral membrane.(ABSTRACT TRUNCATED AT 250 WORDS)