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Thyroid hormone metabolite 3, 5-diiodothyronine (T2) has been shown to possess physiological actions in vertebrates including fishes. It is, however, not certain if T2 has a role in cation transport in fish hepatocytes, particularly in a stressed condition. We, therefore, tested the in vitro action of T2 on the activities of ion transporters such as Na+/K+ ATPase, H+/ K+ ATPase, Na+/NH4 + ATPase, vacuolar H+-ATPase, Plasma Membrane Ca2+ ATPase (PMCA), mitochondrial Ca2+ and mitochondrial H+-ATPase as these ATPases are known for their roles in maintaining systemic and cellular cation gradients including proton and potassium gradients. Hepatocyte explants of air-breathing fish (Anabas testudineus, Bloch), either in non-stressed or hypoxic condition, were incubated with varied doses of T2 (10-9, 10-8 and 10-7 M) for 15 min and the specific activities of these cation-dependent ATPases were analyzed. We found that T2 exposure evoked higher sensitivity to vacuolar and mitochondrial H+-ATPases and H+/K+ ATPase and not to PMCA or mitochondrial Ca2+ ATPase. The data also indicated that T2 has a similar sensitivity to vacuolar and mitochondrial H+-ATPases and H+/K+ ATPase in the hepatocytes of both non-stressed and hypoxia-stressed fish. The data thus provide evidence for a direct action of T2 on the regulation of proton and potassium gradients in the hepatocytes of both non-stressed and hypoxicair-breathing fish.

The ability to respond suitably and maintain a steady state after exposure to stressors is an essential dynamic element in maintaining ion homeostasis. Besides the factors linked to the stressor itself, there are aspects intrinsic to the organisms that are pertinent to shape the stress response, such as age, gender and genetics. This study in mice analyses the functional role of mitochondria, which may affect the integrated responses to psychological stress. Mitochondria depend on a series of ion transporters to interface the communication between the cytosol and the site of energy production, which is key to the survival of the organism. Ion transporters, like mCa2+ATPase, F1F0ATPase and mH+ATPase, are the functional components of the mitochondria involved in Ca2+, H+ homeostasis and energy production. Since the process of aging starts with the birth, and ends with the death of an organism, physiological and molecular processes tend to vary throughout aging. Moreover, males and females have qualitatively different mitochondria, and only a little is known about the mitochondrial responses to stressors. Therefore, we hypothesized that mitochondrial ion transporter functions would modulate the organism’s multisystemic response to psychological stress in an age-, gender- and tissue-specific manner. In this study, BALB/c mice of different age groups (4 weeks-, 8 weeks-, 16 weeks- and 24 weeks-old mice) were subjected to restraint stress of 30 minutes for two consecutive days and the ion transporter activity was quantified in the different regions of the brain (cerebrum, cerebellum and hippocampus) and the gut (duodenum of the intestine, fundus and pyloric regions of the stomach). Overall, the data indicate that in mice both gender-specific and age-specific differential sensitivities to restraint stress exist in mitochondrial ion transporter function in the brain and gut regions. This further points to a decisive interactive role of stress and sex hormones in the energ

The neuronal circuitries of brain and the corresponding ion transporters contribute to the maintenance of ion homeostasis in fish brain. The sensitivity of these neuronal clusters in response to environmental clues brings neural plasticity and subsequent regulation of stress acclimation. Nitric oxide (NO), a gasotransmitter, is involved in ion transport in many peripheral tissues of fishes including air-breathing fish. However, the role of NO in mitochondrial ion transporter activity has not yet been investigated in fish brain. We, therefore, investigated the short-term in vivo action of a NO donor, Sodium Nitro-Prusside (SNP), on mitochondrial ion transporters such as H+- Ca2+- and Mg2+-dependent ATPases in brain segments such as Prosen-Cephalon (PC), Mesen-Cephalon (MC) and Meten-Cephalon (MeC) of immersion-stressed Anabas testudineus. Intraperitoneal injection of SNP for 30 min lowered the activities of bafilomycin-sensitive H+ATPase and vanadate sensitive Ca2+ATPase in PC, whereas in MeC, these transporters showed significant rise in activities after SNP treatment. The oligomycin-sensitive Mg2+ATPase activity showed a significant decrease in PC and MC of brain after SNP treatment in non-stressed fish. Induction of stress by water immersion altered the activities of these ion transporter activities. However, the treatment of SNP in immersed fish showed recovery of the immersion-induced modulation in the activities of these mitochondrial ion transporters. Our data, thus, provide evidence for a decisive role of NO in the recovery process of mitochondrial ion transporters function during immersion stress, confirming a direct differential role of NO in mitochondrial ion homeostasis in teleost brain.

Fishes have evolved complex and multi-step physiological mechanisms to drive ion homeostasis in challenging environments. Induction of stress that disturbs ion homeostasis in fishes evokes recovery response for their survival. Nitric Oxide (NO) as gasotransmitter modulates many physiological mechanisms including ion transport in osmoregulatory epithelia in teleosts. However, little is known about the role of NO in the transport of H+ ions that creates proton gradient with the help of H+-dependent ATPases like H+-ATPase and H+/K+-ATPase, particularly in hepatic and cardiac tissues of bony fish. We, thus, quantified H+-ATPase and H+/K+-ATPase in these tissues after in vivo treatments of NO donor, Sodium Nitro-Prusside (SNP) or NOS inhibitor, L-NAME, in both non-stressed and immersion-stressed air-breathing fish, Anabas testudineus Bloch. We found that elevated NO availability by SNP treatment lowered H+-ATPase-driven H+ transport in both hepatic and cardiac tissues of immersion-stressed fish. In contrast, NO depletion by L-NAME treatment elevated H+-ATPase activity in these tissues of stressed fish, pointing to a direct role of H+-ATPase in NO-mediated proton gradient regulation during stress condition. H+/K+-ATPase that drives H+ transport against K+ reduced its activity in cardiac tissue by SNP and L-NAME treatments. But L-NAME treatment in stressed fish imposed a higher H+ transport in cardiac tissue of these fish. Overall, the data indicate that NO has a vital role in the regulation of H+-ATPase-driven proton gradient in both cardiac and hepatic tissues of immersion-stressed fish.

Thyroid hormones (TH) have a multitude of actions, mainly on development and differentiation during early life and play many vital roles in almost all tissues including neuronal tissues. TH rapidly alters the mitochondrial functions both by its genomic and direct actions on mitochondrial binding sites. The functional relationship between TH and mitochondrial ion transport during stress response has not yet been elucidated in mammals so far. Here, we report a rapid in vivo action of triiodothyronine (T3) on mitochondrial ion transporter functions in the neuronal clusters of cortex, hippocampus and cerebellum of Swiss Albino mouse (Mus musculus) treated short-term with triiodothyronine (T3; 200ng g-1) for 30 min either in non-stressed or in restraint-stressed (30 min each day for 7 days). The mH+-ATPase activity in the cortex decreased to significant levels after T3 treatment in both non-stressed and restraint-stressed mice. On the contrary, the mH+-ATPase activity in the hippocampus and cerebellum increased to significant levels after T3 treatment in both non-stressed and restraint-stressed mice. The mCa2+-ATPase activity in the cortex and cerebellum decreased to significant levels after T3 treatment in both non-stressed and restraint-stressed mice. The mCa2+-ATPase activity in the hippocampus that increased to significant levels after T3 treatment, showed a reversal after restraint-stress in T3-treated mice. The mitochondrial Mg2+-ATPase activity in the cortex decreased to significant levels after T3 treatment in restraint-stressed mice. On the contrary, T3 treatment in restraint stressed mice increased to significant levels the mitochondrial Mg2+-ATPase activity in the cerebellum. The mitochondrial Mg2+-ATPase activity in the hippocampus, which increased to significant levels after T3 treatment in non-stressed mice, reversed its activity in T3-treated restraint-stressed mice. Spatial and differential action of T3 on the mitochondrial ion transporters has been found

The ever-increasing use of plant-based pharmaceuticals as alternatives to conventional drugs for disease management demands identification, isolation, and characterization of novel compounds. Despite the potential of plant extracts to mitigate the morbidity of diseases, several active principles are preferred to avoid the interference of other compounds. The promising health benefits of the extracts and isolated compounds of Pterocarpus santalinus in the treatment of diabetes, cardiovascular disease, cancer, and infections have been described. However, such studies on the active principle, namely, santalins, are not reported. In this study, we standardized the isolation of a mixture of santalins A and B from the heartwood of P. santalinus by column chromatography followed by preparative TLC and HPLC. The partially purified santalins were characterized by LC-MS, HR-MS, and 1H NMR analyses. The isolated combination of santalins displayed higher total antioxidant and DPPH free radical scavenging activity in vitro than the crude heartwood extracts. Administration of the mixture of santalins A and B did not exhibit any antihyperglycemic activity in the liver, kidney, and pancreas of alloxan-induced diabetic rats. However, pretreatment of rats with a mixture of santalins at a dose of 1.0 mg/kg body weight prevented alloxan-induced diabetes as indicated by the normal blood glucose levels. Hyperglycemia-associated lipid peroxidation was abrogated in santalin-pretreated rats that did not develop alloxan-induced diabetes. Furthermore, the alterations in catalase, glutathione peroxidase, and glutathione-S-transferase activities in the pancreas of santalinpretreated rats could be responsible for preventing damage to the pancreas and thus non-induction of diabetes following alloxan treatment. Therefore, for the first time, we report the simplified procedure for isolating a mixture of santalins, including their ability to prevent the induction of diabetes in Wistar rats. The outc

Blackleg disease caused by the fungus Leptosphaeria maculans is one of the most economically significant diseases of canola (Brassica napus L). Three approaches that can be used in mitigating this devastating disease were investigated in this study. Introducing new resistant genotypes is pivotal as host resistance plays the leading role in managing the disease and allows gene rotation. Since AvrLm7 is one of the most abundant avirulence genes in the pathogen population and Rlm7 has been reported as a stable host genotype in other countries, this study focused on evaluating the potential of B. napus genotype, ‘01-23-2-1’, which carries Rlm7. ‘01-23-2-1’was inoculated with L. maculans isolate, carrying AvrLm7, and the CRISPR/Cas9 AvrLm7-knockout mutant of the same isolate to make incompatible and compatible interactions, respectively. A dual RNA-seq study explored differential gene expressions in both interactions. An upregulation of defense-related genes was observed in the incompatible interaction, and in contrast, the same gene expressions were started late in compatible interactions. High expressions of virulence-related pathogen genes-CAZymes, merops, and effector proteins after 7-dpi in compatible interactions but not in incompatible interaction—confirmed that the pathogen was actively virulent only in compatible interactions. Pydiflumetofen is a broad-spectrum seed treatment fungicide in the succinate dehydrogenase inhibitor (SDHI) group. Pydiflumetofen was evaluated compared to Vibrance® Flexi (current company recommendation) under greenhouse conditions using moderately resistant canola cultivar SY4135, and susceptible cultivar Westar. Both SY4135 and Westar plants developed from pydiflumetofen treated seeds showed significantly lower disease severity in canker formation under controlled conditions. Moreover, both host genotypes exhibited lower disease severity in Green fluorescent protein (GFP) tagged L. maculans inoculated seedlings demonstrated control of m

Matrix metalloproteinases 2 and 9 (MMP2 and MMP9) are involved in the extracellular matrix (ECM) remodeling. We tested the short-term in vitro action of inhibitors of MMP2 and MMP9 on P-type ion transporter function in organ explants of climbing perch (Anabas testudineus) to understand how these ECM remodeling components influence the ion transporter function in the osmoregulatory epithelia of fish. Graded doses (10-8, 10-7 and 10-6 M) of inhibitors of MMP2 and MMP9 were administered in vitro to explants of gills, kidney and intestine, kept for either 15 or 30 min and the activities of P-type ATPase such as Na+/K+-ATPase (NKA), H+/K+-ATPase (HKA) and plasma membrane Ca2+-ATPase (PMCA) were quantified. We found that the inhibitors of MMP2 and MMP9 produced dose- and time-dependent modulation in the activities of NKA, HKA and PMCA in the tested tissue explants. Incubation of MMP2 and 9 inhibitors at the highest dose (10-6 M) for 15 and 30 min produced substantial rise in NKA activity. Likewise, HKA activity that showed significant rise after incubation of 10-7 and 10-8 M inhibitors in gills and kidney explants, decreased at the lowest dose (10-8 M) of inhibitors. The lower doses of both inhibitors, while increasing PMCA activity in kidney and intestinal explants inhibited its activity in gill explant. These differential tissue-responsive actions of MMP2 and MMP9 inhibitors indicate that these ECM remodeling components can modify the function of the membrane-bound P-type ion transporters in the osmoregulatory tissues of fish.

Nitric oxide (NO), a gaseous free radical that functions as signal molecule, regulates several physiological functions. But in teleost fishes, the influence of NO on mitochondrial energetics is not yet understood. With a view to understanding the short-term in vivo action of NO on mitochondrial energetics in fish, we examined the effects of sodium nitroprusside, a NO donor (SNP) and N-omega-nitro-L-arginine methyl ester (L-NAME), a competitive inhibitor of nitric oxide synthase (L-NAME), on major electron carriers and oxidative status in heart and liver mitochondria of an obligate air-breathing fish (Anabas testudineus Bloch) kept at either non-stressed or hypoxia-stressed condition. The total nitrate/nitrite (NO3-/NO2-) level that corresponds to NO content showed a rise after SNP (5 μg g-1) and a decline in the heart and liver of non-stressed fish after L-NAME (100 ng g-1) treatments for 30 min. Water immersion for 30 min that induced hypoxia lowered NO3-/NO2- level in heart and liver, but showed a rise in NO3-/NO2- level after SNP treatment of immersion-stressed fish. Reactive Oxygen Species (ROS) production increased after SNP treatment but decreased after L-NAME treatment in heart of hypoxia-stressed fish where as in liver both SNP and L-NAME treatments caused decrease of ROS in stressed fish. SNP treatment increased and L-NAME treatment lowered peroxynitrite (ONOO-) level in heart and liver of non-stressed fish. SNP treatment lowered the activity of cytochrome c oxidase (COX) but L-NAME treatment increased its activity in mitochondria of heart of hypoxia-stressed fish. In liver mitochondria, however, COX activity showed a rise after these treatments. On the contrary, SNP and L-NAME treatments in stressed fish elevated succinate dehydrogenase (SDH) activity in both heart and liver mitochondria. In heart, LDH activity increased after SNP and L-NAME treatments in both non-stressed and stressed conditions, but not in liver of stressed fish. Put together, the data

Purification and Characterization of LTC4 Synthase from Sheep UterusPurification and Characterization of LTC4 Synthase from Sheep UterusEicosanoids, the oxygenated metabolites of eicosapolyenoic fatty acids such as arachidonic acid via the cyclooxygenase (COX), lipoxygenase (LOX) and epoxygenase (EPOX) pathways, are generated in response to specific stimuli, elicit the response and are then quickly metabolized. Hence, these are rightly termed as “local hormones” or “autocoids”. They are involved in the regulation of a variety of physiological as well as pathological processes, including reproduction. While there are extensive studies on the role of COX metabolites, such as prostaglandins, in reproduction, not much is known on the role of LOX metabolites in reproduction. Earlier, we have identified abundant LOX activity in sheep uterus and the highly purified enzyme was found to be a homo-dimeric protein with a molecular weight of 66 kDa. When incubated with arachidonic acid, the enzyme showed two lipoxygenase activities producing both 12- and 15-Hydroxyeicosatetraenoic acid (15-HPETEs) at the optimum pH of 5.5. The relative concentration of 12- and 15-HPETEs, however, changed with the pH of the reaction, 12-Hydroxyeicosatetraenoic acid (HETE) being higher in the alkaline range and 15-HETE being the abundant in the acidic range. Furthermore, the enzyme showed the dual lipoxygenase based 14,15-LTA4 synthase activity as evidenced by the formation of 8,15-diHETEs, the hydrolysis products of 14,15-LTA4. In the present study, leukotriene C4 synthase (LTC4S) enzyme was purified on Q-Sepharose column after solubilization of microsomes utilizing a combination of CHAPS and taurocholate. The purified enzyme showed activity with 5, 6-LTA4 and 14, 15-LTA4, with slight preference towards the latter, and converting them to corresponding LTC4s. Both methyl esters and free acids of LTA4 served as substrates, though the activity was more with methyl esters. However, the enzym

The morphometric analysis was carried out in indigenous cyprinid food fish, Schizothorax labiatus (a species of snow trout) collected from Vishav stream, a perennial cold water stream of Kashmir valley, India, wherein various morphometric characters viz., Total Length (TL), Standard Length (SL), Fork Length (FL), Pre-dorsal Length (PDL), Pre-pectoral Length (PPL), Pre-pelvic Length (PPeL), Pre-anal Length (PAL), Anal Fin Length (AL), Head Length (HL), Height of Dorsal Fin (HOD), Snout Length (SnL), Body Depth (BD), Eye Diameter (ED), Inter Orbital Length (IOL), Caudal Fin Length (CL) and Caudal Depth (CD) of 104 fish specimens were measured, correlated and analysed. The morphometric data was calculated in the percentage of total length except SnL, ED and IOL which were calculated in the percentage of HL. The result revealed that the correlation coefficient ranged from 0.770 to 0.995 in proportion to TL and 0.706 to 0.859 in proportion to HL which indicates that all the morphometric characters are highly correlated. All the correlations were significant at the level of p<0.01. During the investigation, the results revealed that in the % of TL, 8 characters were found to be genetically controlled 4 characters as intermediate and none character as environmentally controlled one. In the % of HL, 1 character was found to be included in intermediate character, 2 characters in environmentally controlled characters and none in purely genetically controlled one. As revealed by the regression analysis of different morphometric characters in proportion to TL, the maximum growth was obtained for FL (b = 0.9664) and the minimum growth was obtained for AL (b = 0.0655) and in proportion to HL, the maximum growth was obtained for IOL (b = 0.4689) and the minimum growth was obtained for ED (b = 0.0852).

Ammonia, as an endogenous respiratory gas, is produced during protein and amino acid metabolism. Accumulation of excess ammonia is toxic, and fishes have developed mechanisms to defend against ammonia toxicity. Here, we tested the in vivo action of ammonia in an air-breathing fish to find how it modulates mitochondrial ion transport in fish heart and liver. We thus analysed the activity pattern of mitochondrial ion transporters such as mitochondrial Ca2+ATPase, mitochondrial H+ATPase and mitochondrial F1F0 ATPase in heart and liver of air-breathing fish Anabas testudineus which were kept for immersion-induced hypoxia stress. In addition, plasma metabolites such as glucose and lactate were also quantified. Oral administration of ammonia solution [(NH4)2SO4; 50ng g-1] for  30 min increased mit.Ca2+ATPase activity in heart but lowered its activity in liver mitochondria. A reduced mit.H+ATPase activity was found in heart but in liver its activity showed increase after ammonia treatment. F1F0 ATPase increased significantly in heart but showed reduced activity in liver mitochondria. Administration of ammonia in immersion-stressed fish, however, nullified the excitatory response of heart and liver mitochondria in the immersion-stressed fish. Overall, the data indicate that ammonia can play a significant physiological role in the regulation of mitochondrial ion homeostasis in the liver and heart of air-breathing fish during their acclimation to hypoxia stress.

type:Journal Article, Recent studies demonstrated that the corticospinal pathway is one of the key nodes for the feedback control of human standing and that the excitability is flexibly changed according to the current state of posture. However, it has been unclear whether this pathway is also involved in a predictive control of human standing. Here, we investigated whether the corticospinal excitability of the soleus (SOL) and tibialis anterior (TA) muscles during standing would be modulated anticipatorily when perturbation was impending. We measured the motor-evoked potential (MEP) induced by transcranial magnetic stimulation over the motor cortex at six stimulus intensities. Three experimental conditions were set depending on predictabilities about perturbation occurrence and onset: No perturbation, No Cue, and Cue conditions. In the Cue condition, an acoustic signal was given as timing information of perturbation. The slope of the stimulus–response relation curve revealed that the TA-MEP was enhanced when postural perturbation was expected compared to when the perturbation was not expected (No Perturbation vs. No Cue, 0.023 ± 0.004 vs. 0.042 ± 0.007; No Perturbation vs. Cue, 0.023 ± 0.004 vs. 0.050 ± 0.009; Bonferroni correction, p = 0.01, respectively). In addition, two-way analysis of variance (intensity × condition) revealed the main effect of condition (F(1,13) = 6.31, p = 0.03) but not intensity and interaction when the MEP amplitude of the Cue and No Cue conditions was normalized by that in No Perturbation, suggesting the enhancement more apparent when timing information was given. The SOL-MEP was not modulated even when perturbation was expected, but it slightly reduced due to the timing information. The results of an additional experiment confirmed that the acoustic cue by itself did not affect the TA- and SOL-MEPs. Our findings suggest that a prediction of a future state of standing balance modulates the corticospinal excitability in the TA, and that the additional timing in, source:https://doi.org/10.3389/fnhum.2018.00068

type:Journal Article, The purpose of the present study was to investigate how stretch reflex (SR) responses in the ankle extensor (soleus: SOL) and flexor (tibialis anterior: TA) muscles would be modulated with temporal and/or spatial predictions of external perturbations and whether their effects are specific to the standing posture. SR responses in the SOL/TA were elicited by imposing quick ankle toes-up/toes-down rotations while standing upright and in the supine position. We designed four experimental conditions based on pre-information about perturbations: no information (No Cue), the timing of the perturbation onset (TIM), the direction of the perturbation (DIR), and both the timing and direction of the perturbation (TIM/DIR). Each condition was separated and its order was counterbalanced. In the SR of TA evoked by toes-down rotation, integrated electromyography activities of the late component were significantly reduced in the TIM and TIM/DIR conditions as compared with those in the No Cue and DIR conditions. The occurrence rate of late SR components that reflects how often the reflex response was observed was also lower in the TIM and TIM/DIR conditions as compared with that in the No Cue and DIR conditions. On the other hand, no significant changes were seen among the four conditions in the early SR component in the TA and both SR components in the SOL. The same results in the occurrence rate were found in the supine position. The present results suggest (1) only temporal predictions have a remarkable effect on the SR excitability of the TA, and (2) this effect is independent of posture., source:https://doi.org/10.1371/journal.pone.0158721

type:Journal Article, The prediction of upcoming perturbation modulates postural responses in the ankle muscles. The effects of this prediction on postural responses vary according to predictable factors. When the amplitude of perturbation can be predicted, the long-latency response is set at an appropriate size for the required response, whereas when the direction of perturbation can be predicted, there is no effect. The neural mechanisms underlying these phenomena are poorly understood. Here, we examined how the corticospinal excitability of the ankle muscles [i.e., the tibialis anterior (TA), the soleus (SOL), and the medial gastrocnemius (MG), with a focus on the TA], would be modulated in five experimental conditions: (1) No-perturbation; (2) Low (anterior translation with small amplitude); (3) High (anterior translation with large amplitude); (4) Posterior (posterior translation with large amplitude); and (5) Random (Low, High, and Posterior in randomized order). We measured the motor-evoked potentials (MEPs) induced by transcranial magnetic stimulation (TMS) at 50 ms before surface-translation in each condition. The electromyographic (EMG) responses evoked by surface-translations were also measured. The results showed that the TA-MEP amplitude was greater in the High condition (where the largest TA-EMG response was evoked among the five conditions) compared to that in the No-perturbation, Low, and Posterior conditions (High vs. No-perturbation, p < 0.001; High vs. Low, p = 0.001; High vs. Posterior, p = 0.001). In addition, the MEP amplitude in the Random condition was significantly greater than that in the No-perturbation and Low conditions (Random vs. No-perturbation, p = 0.002; Random vs. Low, p = 0.002). The EMG response in the TA evoked by perturbation was significantly smaller when a perturbation can be predicted (predictable vs. unpredictable, p < 0.001). In the SOL and MG muscles, no prominent modulations of the MEP amplitude or EMG response were observed, suggesting that, source:https://doi.org/10.3389/fnhum.2019.00004

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Photosynthetic bacteria offer excellent experimental opportunities to explore both the structure and function of the ubiquinol-cytochrome c oxidoreductase (bc 1 complex). In both Rhodobacter sphaeroides and Rhodobacter capsulatus, the bc 1 complex functions in both the aerobic respiratory chain and as an essential component of the photosynthetic electron transport chain. Because the bc 1 complex in these organisms can be functionally coupled to the photosynthetic reaction center, flash photolysis can be used to study electron flow through the enzyme and to examine the effects of various amino acid substitutions. During the past several years, numerous mutations have been generated in the cytochrome b subunit, in the Rieske iron-sulfur subunit, and in the cytochrome c 1 subunit. Both site-directed and random mutagenesis procedures have been utilized. Studies of these mutations have identified amino acid residues that are metal ligands, as well as those residues that are at or near either the quinol oxidase (Q o) site or the quinol reductase (Q i) site. The postulate that these two Q-sites are located on opposite sides of the membrane is supported by these studies. Current research is directed at exploring the details of the catalytic mechanism, the nature of the subunit interactions, and the assembly of this enzyme.

Mutation, irregular expression and sustained activation of TRPM4 have been linked to various cardiovascular diseases, including ischemia-reperfusion injury, stroke, arrhythmia (6% Brugada cases), cardiac conduction disorders and hypertension. However, to date much remains unknown about its structure. Here a heterologously expressed human TRPM4-GFP fusion protein was purified, then its supramolecular structure and functionality was examined. The oligomeric state of TRPM4-GFP in detergent micelles was characterised using number and brightness analysis, crosslinking studies, native gel electrophoresis, multi-angle laser light scattering and electron microscopy. Our data demonstrate that TRPM4 is tetrameric, which is the case for other TRP channels studied to date. To assess the functionality of the protein we have obtained single-channel recordings from hTRPM4-eGFP proteoliposomes, which demonstrate inhibition by Flufenamic acid (FFA), suggesting that the channels are fully functional upon reconstitution. The fact that purified TRPM4-GFP maintains a tetrameric form in detergent and exhibits characteristic channel activity indicates that the channel protein we have produced is functional. This body of work will enable future more detailed examination of the structural and functional properties of this therapeutically significant channel.

http://www.tulips.tsukuba.ac.jp/mylimedio/dl/page.do?issueid=103000249&old_issueid=1106965&tocid=100098528&page=9-18

The study aims to evaluate the wound healing properties of bioactive fractions from the extract of Butea monosperma (Lam) stem bark. In this study the stem bark powder was extracted with ethanol, further the ethanolic extract was fractionated with different solvents (petroleum ether, benzene, chloroform and acetone) in increasing order of polarity. Thus prepared extracts were subjected to preliminary phytochemical analysis. The wound healing activity of the ethanolic extract and the fractions isolated from the stem bark of Butea monosperma were evaluated in excision, incision and dead space wound healing models using Albino wistar rats. The wound healing activity was assessed by the breaking strength in case of incision wounds, epithelialization and wound contraction in case of excision wound and granulation tissue dry weight, breaking strength and hydroxyproline content in case of dead space wound. The ethanolic extract and the acetone fraction showed the significant wound healing activity on all three wound models. The phytochemical investigations revealed the presence of alkaloids, tannins, flavonoids, phenolic compounds and steroids. The increased rate of wound contraction and hydroxyproline content in the ethanolic extract and the acetone fraction treated animals provides a scientific base to the ethno medicinal use of Butea monosperma, which is largely attributable to the additive or synergistic effect of their constituents. Keywords: Butea monosperma, Dead space wound, Excision wound, Incision wound.

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Neste estudo foram realizados bioensaios para avaliar o possível efeito atrativo e/ou estimulante de extratos de ovos de Lutzomyia renei. A resposta ao feromônio de oviposição pela L. renei não foi detectada como para L. longipalpis and Phlebotomus papatasi. Extratos equivalentes a 100 ou 200 ovos não produziram atração perceptível e/ou qualquer estimulo à oviposição, embora extratos de 1000 ovos produziram um leve estimulo atrativo. Análise química dos extratos de ovos de L. renei revelaram a presença de vários ácidos graxos e experimentos complementario são necessários para provar o possível efeito estimulante., In this study bioassays were carried out to evaluate the possible attractive and/or stimulant effect of extracts of the eggs of gravid female Lutzomyia renei. The response to the oviposition pheromone by L. renei was not as marked as that found for L. longipalpis and Phlebotomus papatasi. Extracts equivalent to 100 or 200 eggs did not produce a perceptible attraction and/or stimulation to oviposit, although extracts of 1000 eggs did produce a slight attractive response. Chemical analysis of L. renei egg extracts revealed the presence of various fatty acids and complementary bioassay experiments are needed to prove a possible stimulant effect.

The order of eight of the 29 endonuclease EcoRI-generated fragments of chloroplast DNA was determined. Three sets of rRNA genes aligned sequentially in the same orientation form part of this region. The repeated sets differ in the length and sequence of the spacers among themselves and with the rRNA genes of E. gracilis strain Z.

The nerve fibre loss, atrophy and injury of diabetic peripheral polyneuropathy and their responses to metabolic intervention have been studied by morphometric analysis of sural nerve biopsies. The magnitudes and sources of intra- and inter-individual variation in these morphometric measures have not been investigated previously in a systematic manner. Morphometric parameters of nerve fibre damage were measured in four separate fascicles from bilateral sural nerve specimens obtained post-mortem from 13 diabetic and 13 non-diabetic subjects. Intra- and inter-individual coefficients of variation were computed and compared to the magnitude of the differences between normal and diabetic subjects. Several morphometric variables emerged as highly sensitive and reproducible measures of nerve fibre damage suitable for clinical studies of diabetic peripheral polyneuropathy. These observations provide a rational basis for the design of future clinical trials employing morphometric end-points.

Casein messenger RNAs (mRNAcsn) were purified from lactating mammary glands of BALB/c mice and used as a starting material for cloning of casein gene sequences. Double-stranded casein cDNA (ds-cDNAscn) was prepared and blunt-end ligated to HindIII-specific DNA linker molecules. After digestion with HindIII, the dsDNAcsn was inserted into the HindIII site of plasmid pBR322, using T4 DNA ligase. Escherchia coli strain RH202 was transformed with the hybrid plasmids, and transformants were selected for resistance to ampicillin. Electrophoresis of HindIII-digested hybrid plasmid DNAs, followed by Souther transfer and hybridization to 9[32P]cDNAcsn, revealed that one of the hybrid-plasmid-containing colonies, designated pCas51, contained a 400-bp insert which hybridized to the [32P]cDNAcsn. Purification of the individual casein mRNAs (mRNAcsn [alpha], [beta] and [gamma]) and solution hybridization of nick-translated insert DNA to each of these revealed that pCas51 contained sequences complementary primarily to mRNAcsn [beta].

http://www.tulips.tsukuba.ac.jp/mylimedio/dl/page.do?issueid=102758242&old_issueid=847405&tocid=100062068&page=1-19

本研究では、筑波大学農林技術センター圃場において、2003年にサツマイモ栽培における素材の異なるマルチ資材6種類を用いた要因実験を実施して、資材特性とサツマイモの生育特性、収量性との関係を検討した。被覆資材としてポリエチレンフィルムでは透明(PE-1)および黒(PE-2)の2種、生分解性フィルムでは、ポリブチルサクシネート系フィルム2種(PBS-1、2)、高分子系デンプンフィルム1種(HMS-1)、オレフィン分解付与系フィルム1種(POF-1)の計6種を使用した。地温の推移は、いずれの被覆資材の実験区も同様な動きを示したが、積算地温ではPE-1区が最も高かった。また、土壌水分含有率はPE-1,PE-2,PBS-1,POF-1の4区が高く推移した。初期成育はPOF-1区が最も良好であった。収量性は、塊根数ではPE-1区とPOF-1区は同等であったが、塊根重ではPOF-1、HMS-1区がPE-1区よりも勝っていた。以上の結果から、生分解性マルチ資材では、資材特性として地温保持能力が高く、崩壊開始が遅いPOF-1, HMS-1が望ましいと考えられた。, http://www.tulips.tsukuba.ac.jp/mylimedio/dl/page.do?issueid=102713411&old_issueid=799645&tocid=100052158&page=1-13

"Under contract number: DACW25-79-C-0008."; "Final draft of a report to the Great River Environmental Action Team (Great II) and the Rock Island District, U.S. Army Corps of Engineers.", Item donated by the Upper Mississippi River Conservation Committee (UMRCC) to Murphy Library in 2018. Part of the UMRCC Collection housed at Murphy Library, University of Wisconsin-La Crosse.

The magnetic properties (first-order reversal curves, ferromagnetic resonance and decomposition of saturation remanent magnetization acquisition) of Magnetovibrio blakemorei, a cultivated marine magnetotactic bacterium, differ from those of other magnetotactic species from sediments deposited in lakes and marine habitats previously studied. This finding suggests that magnetite produced by some magnetotactic bacteria retains magnetic properties in relation to the crystallographic structure of the magnetic phase produced and thus might represent a ‘magnetic fingerprint’ for a specific magnetotactic bacterium. The use of this fingerprint is a non-destructive, new technology that might allow for the identification and presence of specific species or types of magnetotactic bacteria in certain environments such as sediments.