An Effect of pH and Bicarbonate on Salt Accumulation by Disks of Storage Tissue

1. The uptake of potassium chloride by disks of several storagetissues has been determined from solutions at alkaline pH valuesand compared with that form solutions at pH 6.5. The stimulationof cation uptake under the alkaline conditions which has previouslybeen obtained with beet tissues was observed with some of thesetissues to a greater or lesser extent. 2. It is concluded that it is the accumulation of anions present in alkaline solutions which generally promotesthe uptake of cations; tissues which do not show the ‘pHeffect’ accumulate very little in the presence of the Cl^- anion. 3. In some cases the alkaline pH values also stimulated anionuptake. This was especially true in artichoke in which the absorptionof both ions was stimulated by approximately the same amount.It is concluded that this represents either a true pH effector a direct effect of anions on the overall salt sccumulation mechanism rather than an indirect effect throughorganic acid synthesis. 4. The experiments reveal a cation/anion ratio of approximatelyI:I when the uptake of ions is taken into consideration. It is suggested that this ratio may be more frequent then isgenerally accepted and that the uptake of CO₂ (converted toan anion in the tissue) may take the place ions at lower pH values. 5. The deleterious effects of frequently observed on growth contrast with the stimulations of salt absorptionobtained here; the rapid increase in the concertration of cationsand/or organic acid anions associated with HCO uptake may insome way be responsible for this.     

Bicarbonate and fast-twitch muscle: Evidence for a major role in pH regulation

Summary Internal pH (pH_i) was analyzed in rat extensor digitorum longus (Edl) muscle at 30°C with single-barrel liquid ionselective electrodes. Average pH_i in 284 cells was 7.197±0.006. Increases in CO₂ from nominally 0 to 5% produced an acidification from which recovery took place. In different groups of cells, recovery from the 5% CO₂ acidification was significantly inhibited by 100 m 4,4 diisothiocyanatostilbene 2,2 disulfonic acid (DIDS), Cl removal, Na removal and 2mm amiloride. Prepulsing with 20mm NH₄ in the presence of CO₂/HCO₃ typically reduced pH_i to only about neutral, whereas 50mm reduced pH_i to 6.7–6.8. In the nominal absence of CO₂/HCO₃, 20mm NH₄ reduced pH_i to about 6.7 from which recovery took place at about 58% of the rate seen in different cells in the presence of CO₂/HCO₃. In the presence of CO₂/HCO₃, cells prepulsed with 50mm NH₄ had fully recovered to an average pH_i of 7.22±0.04 about 90 min after removal of NH₄. However, 90 min after removal of 20mm NH₄ in the absence of CO₂/HCO₃, average pH_i was significantly less (7.05±0.03). Intrinsic buffering capacity ( _i ) was obtained during pulses of CO₂, acetic acid or after an NH₄ pulse, _i was significantly reduced in the absence of HCO₃, Cl or Na and HCO₃. The data provide significant support for an important role of HCO₃ in the control of pH_i in fast-twitch muscle.     

An Automated System for Rapid Non-Destructive Enumeration of Growing Microbes

Background

The power and simplicity of visual colony counting have made it the mainstay of microbiological analysis for more than 130 years. A disadvantage of the method is the long time required to generate visible colonies from cells in a sample. New rapid testing technologies generally have failed to maintain one or more of the major advantages of culture-based methods.

Principal Findings

We present a new technology and platform that uses digital imaging of cellular autofluorescence to detect and enumerate growing microcolonies many generations before they become visible to the eye. The data presented demonstrate that the method preserves the viability of the microcolonies it detects, thus enabling generation of pure cultures for microbial identification. While visual colony counting detects Escherichia coli colonies containing about 5×10⁶ cells, the new imaging method detects E. coli microcolonies when they contain about 120 cells and microcolonies of the yeast Candida albicans when they contain only about 12 cells. We demonstrate that digital imaging of microcolony autofluorescence detects a broad spectrum of prokaryotic and eukaryotic microbes and present a model for predicting the time to detection for individual strains. Results from the analysis of environmental samples from pharmaceutical manufacturing plants containing a mixture of unidentified microbes demonstrate the method''s improved test turnaround times.

Conclusion

This work demonstrates a new technology and automated platform that substantially shortens test times while maintaining key advantages of the current methods.     

Bicarbonate determines cytoplasmic pH and suppresses mitogen-induced alkalinization in fibroblastic cells

Addition of growth factors to responsive cells in HCO3- -free media results in a rapid rise in cytoplasmic pH (pHi) caused by activation of Na+/H+ exchange. In this paper, we have examined how pHi regulation and growth factor responsiveness are affected by HCO3(-)using quiescent mouse MES-1 fibroblastic cells as a model. When cells are exposed to 25 mM HCO3-, 5% CO2, steady-state pHi reaches a new more alkaline level (by 0.25 unit) within 10 min. This rise in pHi is both Na+- and HCO3- -dependent, does not occur in Cl(-)-depleted cells, and is inhibited by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid, but not by 5-(n,n-dimethyl)-amiloride, indicating the involvement of Na+-dependent HCO3-/Cl- exchange. Furthermore, the recovery of pHi from acute acid loads is accelerated by HCO3- in a Na+-dependent and 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid-sensitive manner and is blocked in Cl(-) -depleted cells. Similar results were obtained for mouse 3T3 cells and human fibroblasts. In the presence of HCO3-/CO2 (pH 7.35), mitogens and phorbol esters fail to induce a detectable rise in pHi. However, when steady-state pHi is artificially lowered by approximately 0.4 unit, growth factors evoke significant increases in pHi due to activation of Na+/H+ exchange. In the absence of HCO3-, mitogen-induced alkalinizations are readily detectable but not when pHi is artificially elevated to the value normally observed in HCO3- media. From these results we conclude that: 1) Na+-dependent HCO3-/Cl- exchange determines steady-state pHi and acts in parallel with Na+/H+ exchange to stimulate pHi recovery from acid loading; 2) Na+-dependent HCO3-/Cl- exchange raises steady-state pHi to a level beyond the operating range of the Na+/H+ exchanger and thereby prevents growth factors from alkalinizing the cytoplasm any further. The results also imply that, unlike Na+/H+ exchange, Na+-dependent HCO3-/Cl- exchange is not activated by mitogens.     

An Automated System for Skeletal Maturity Assessment by Extreme Learning Machines

Assessing skeletal age is a subjective and tedious examination process. Hence, automated assessment methods have been developed to replace manual evaluation in medical applications. In this study, a new fully automated method based on content-based image retrieval and using extreme learning machines (ELM) is designed and adapted to assess skeletal maturity. The main novelty of this approach is it overcomes the segmentation problem as suffered by existing systems. The estimation results of ELM models are compared with those of genetic programming (GP) and artificial neural networks (ANNs) models. The experimental results signify improvement in assessment accuracy over GP and ANN, while generalization capability is possible with the ELM approach. Moreover, the results are indicated that the ELM model developed can be used confidently in further work on formulating novel models of skeletal age assessment strategies. According to the experimental results, the new presented method has the capacity to learn many hundreds of times faster than traditional learning methods and it has sufficient overall performance in many aspects. It has conclusively been found that applying ELM is particularly promising as an alternative method for evaluating skeletal age.     

A Permanent Automated Real-Time Passive Acoustic Monitoring System for Bottlenose Dolphin Conservation in the Mediterranean Sea

Within the framework of the EU Life+ project named LIFE09 NAT/IT/000190 ARION, a permanent automated real-time passive acoustic monitoring system for the improvement of the conservation status of the transient and resident population of bottlenose dolphin (Tursiops truncatus) has been implemented and installed in the Portofino Marine Protected Area (MPA), Ligurian Sea. The system is able to detect the simultaneous presence of dolphins and boats in the area and to give their position in real time. This information is used to prevent collisions by diffusing warning messages to all the categories involved (tourists, professional fishermen and so on). The system consists of two gps-synchronized acoustic units, based on a particular type of marine buoy (elastic beacon), deployed about 1 km off the Portofino headland. Each one is equipped with a four-hydrophone array and an onboard acquisition system which can record the typical social communication whistles emitted by the dolphins and the sound emitted by boat engines. Signals are pre-filtered, digitized and then broadcast to the ground station via wi-fi. The raw data are elaborated to get the direction of the acoustic target to each unit, and hence the position of dolphins and boats in real time by triangulation.     

An Automated and Intelligent Medical Decision Support System for Brain MRI Scans Classification

A wide interest has been observed in the medical health care applications that interpret neuroimaging scans by machine learning systems. This research proposes an intelligent, automatic, accurate, and robust classification technique to classify the human brain magnetic resonance image (MRI) as normal or abnormal, to cater down the human error during identifying the diseases in brain MRIs. In this study, fast discrete wavelet transform (DWT), principal component analysis (PCA), and least squares support vector machine (LS-SVM) are used as basic components. Firstly, fast DWT is employed to extract the salient features of brain MRI, followed by PCA, which reduces the dimensions of the features. These reduced feature vectors also shrink the memory storage consumption by 99.5%. At last, an advanced classification technique based on LS-SVM is applied to brain MR image classification using reduced features. For improving the efficiency, LS-SVM is used with non-linear radial basis function (RBF) kernel. The proposed algorithm intelligently determines the optimized values of the hyper-parameters of the RBF kernel and also applied k-fold stratified cross validation to enhance the generalization of the system. The method was tested by 340 patients’ benchmark datasets of T1-weighted and T2-weighted scans. From the analysis of experimental results and performance comparisons, it is observed that the proposed medical decision support system outperformed all other modern classifiers and achieves 100% accuracy rate (specificity/sensitivity 100%/100%). Furthermore, in terms of computation time, the proposed technique is significantly faster than the recent well-known methods, and it improves the efficiency by 71%, 3%, and 4% on feature extraction stage, feature reduction stage, and classification stage, respectively. These results indicate that the proposed well-trained machine learning system has the potential to make accurate predictions about brain abnormalities from the individual subjects, therefore, it can be used as a significant tool in clinical practice.     

Ultrasonic Telemetry, Tracking and Automated Monitoring Technology for Sharks

Sharks were among the first marine animals to carry telemetry systems because of their size and the need to understand their interactions with humans. Modern telemetry systems can gather many kinds of data (limited only by imagination, funding and sensor types), indicating which animals are near telemetry receivers and what they are doing. Receivers now range from simple autonomous detector units for deployment in mid-water in large-scale grids, to sophisticated automated benthic recorders, to triangulating radio-linked buoy systems (RAP), to ship-borne transponders. In addition, archival tags can now gather and store data even while the shark is away, to be downloaded later. Older types had to be recovered, but popup tags release from sharks automatically, surface and transfer data to satellites, while CHAT tags download whenever queried by a nearby transponding acoustic receiver. Sophisticated animal-borne tags dramatically increase the information gathered about sharks and their environment. The examples provided show the parallel progression of shark biology and acoustic biotelemetry illustrating that telemetry systems are tools for gathering data, which can often be honed to facilitate biological experiments. Future visions include sensors that directly measure shark swimming power and cardiac output, compressing the data so that it can be delivered to RAP systems tracking multiple animals with meter resolution in near real time. CHAT tags as small as 22mm diameter should be able to return similar data from trips of hundreds of kilometers. Continued communication between biologists and engineers is essential to develop these technologies.     

北京大学第三医院能耗监管系统的构建与运用

近年来,随着我国医院医疗科技的发展和诊疗环境的改善,医院的能源消耗也在不断地上涨。掌握医院的用能模式并对能源数据加以统计分析是开展能源管理的基础。文章基于北京大学第三医院已建成并投入使用的能耗监管系统,通过能耗计量、数据分析、节能诊断和指标管理等措施,积极开展医院能耗的应用研究,扎实有效地推进医院能源管理工作,以期达到提高医院能源的使用效率,优化医院运营管理的目的。     

Buffers for the reconstitution of aspartate aminotransferase

The cofactor activation of the apoenzyme of pig heart cytosolic aspartate aminotransferase was studied in various buffers. Cationic buffers are shown to allow maximal reconstitution in the pH range of 5.0 to 9.0. Anionic buffers made up of mono- and dicarboxylates are found to affect reconstitution in a pH-dependent manner. At low pH, the carboxylates strongly inhibit reconstitution, but at high pH, they show less effect. In contrast, the more potent inhibitor Pi shows the opposite pH profile. Dicarboxylates are considerably more inhibitory than monocarboxylates. Substantial protection against inhibition by a number of carboxylates may be achieved by the addition of sodium chloride.     

The Effect of pH and Bicarbonate Ions on the Uptake of Salts by Disks of Red Beet

1. The uptake of potassium chloride by disks of beetroot in solutionsat different pH values has been followed over periods of severaldays.
2. It was found that, as the pH was raised from approximately6to 9 the rate of uptake of potassium from 0.0024 M. potassiumchloride increased, and there was a corresponding increase inthe amount of potassium held by the disks at satura tion. Boththe rate of uptake and the final content of potassium in diskskept in solutions at pH 8.5 were frequently double those atpH 6.
3. The rate of uptake of chloride ions was only slightlyaffectedby the pH value of the medium. The final chloride contentwasapproximately the same in disks kept at different pH values.
4. The above ‘pH effect’ on potassium uptake hasbeenshown to be related to the uptake of bicarbonate ions presentin the alkaline solution; the additional potassium ions accumulatedaccompanying the bicarbonate anions taken up.
5. The bicarbonateions accumulated are converted primarily tomalic acid.

     

Chronobiology of Aging in Albizzia julibrissin: I. An Automated, Computerized System for Monitoring Leaflet Movement; The Rhythm in Constant Darkness

We are using leaflet movements in Albizzia julibrissin as a model system for investigating the chronobiology of aging. To monitor leaflet movements during long dark periods with adequate temporal and spatial resolution, we designed an automated, computerized photoelectric monitoring system. Each of 12 leaflet pairs was positioned in an individual light-proof container, with one leaflet immobilized. The angle of the mobile leaflet was monitored by a photosensor array using a low intensity infrared beam. Leaflet position was determined by custom-developed software, using information on the shading patterns of the sensors. Data on leaflet angle as a function of time were collected and stored on a floppy disc and then printed in numerical and graphical form.

Oscillations of young, middle-aged, and old leaflets persist during 7 d of darkness with a periodicity close to 24 h. Period length appears to be age-independent, but rhythmic wave form is age-dependent. The older the leaflet, the earlier and more completely it opens and the less completely it closes.

     

Bicarbonate infusion and pH clamp moderately reduce hyperventilation during ramp exercise in humans.

To test the hypothesis that the decrease in plasma pH contributes to the hyperventilation observed in humans in response to exercise at high workloads, five healthy male subjects performed a ramp exercise [maximal workload: 352 W (SD 35)] in a control situation and when arterialized plasma pH was maintained at the resting level (pH clamp) by intravenous infusion of sodium bicarbonate [129 mmol (SD 23), beginning at 59% maximal workload (SD 5)]. Bicarbonate infusion did not modify O(2) consumption (Vo(2)) but significantly (P < 0.05) increased arterial Pco(2), plasma bicarbonate concentration, and respiratory exchange ratio (P < 0.05). At the three highest workloads, pulmonary ventilation (Ve) and Ve/Vo(2) were approximately 5-10% lower (P < 0.05) when bicarbonate was infused than in the control situation, and hyperventilation was reduced by 15-30%. These data suggest that the decrease in plasma pH is one of the factors that contribute to the hyperventilation observed at high workloads.     

The Relationship Between Bicarbonate Ion, pH, and 59Fe Uptake by Plants

Iron absorption by barley, pea, and runner bean is stimulatedby bicarbonate but inhibited by increasing pH. Bicarbonate stimulatestranslocation of Fe in barley but inhibits it in pea and bean.Increasing pH stimulates translocation in barley and pea withno effect in bean. The presence of other ions in the externalsolution appears to inhibit absorption but enhances translocationof Fe in bean and pea. It is concluded that whether or not bicarbonatewill induce chlorosis in a plant depends on species and variety.     

ConfocalCheck - A Software Tool for the Automated Monitoring of Confocal Microscope Performance

Laser scanning confocal microscopy has become an invaluable tool in biomedical research but regular quality testing is vital to maintain the system’s performance for diagnostic and research purposes.Although many methods have been devised over the years to characterise specific aspects of a confocal microscope like measuring the optical point spread function or the field illumination, only very few analysis tools are available. Our aim was to develop a comprehensive quality assurance framework ranging from image acquisition to automated analysis and documentation. We created standardised test data to assess the performance of the lasers, the objective lenses and other key components required for optimum confocal operation.The ConfocalCheck software presented here analyses the data fully automatically. It creates numerous visual outputs indicating potential issues requiring further investigation. By storing results in a web browser compatible file format the software greatly simplifies record keeping allowing the operator to quickly compare old and new data and to spot developing trends.We demonstrate that the systematic monitoring of confocal performance is essential in a core facility environment and how the quantitative measurements obtained can be used for the detailed characterisation of system components as well as for comparisons across multiple instruments.     

Bicarbonate/chloride antiport in Vero cells: II. Mechanisms for bicarbonate-dependent regulation of intracellular pH

The rates of bicarbonate-dependent uptake and efflux of 22Na+ in Vero cells were studied and compared with the uptake and efflux of 36Cl-. Both processes were strongly inhibited by DIDS. Whereas the transport of chloride increased approximately ten-fold when the internal pH was increased over a narrow range around neutrality, the uptake of Na+ was much less affected by changes in pH. The bicarbonate-linked uptake of 22Na+ was dependent on internal Cl- but not on internal Na+. At a constant external concentration of HCO3-, the amount of 22Na+ associated with the cells increased when the internal concentration of HCO3- decreased and vice versa, which is compatible with the possibility that the ion pair NaCO3- is the transported species and that the transport is symmetric across the membrane. Bicarbonate inhibited the uptake of 36Cl- both in the absence and presence of Na+. At alkaline internal pH, HCO3- stimulated the efflux of 36Cl- from preloaded cells, while at acidic internal pH both Na+ and HCO3- were required to induce 36Cl- efflux. We propose a model for how bicarbonate-dependent regulation of the internal pH may occur. This model implies the existence of two bicarbonate transport mechanisms that, under physiological conditions, transport OH(-)-equivalents in opposite directions across the plasma membrane.     

Bicarbonate/chloride exchange regulates intracellular pH of embryos but not oocytes of the hamster.

The ability to regulate intracellular pH (pH(i)) is essential for normal cell development and differentiation. This study was an investigation of the regulatory system used by the hamster oocyte and preimplantation embryo to regulate pH(i) in the alkaline range. Recovery from alkalosis by late 1-cell and 2-cell embryos was rapid, and physiological pH(i) levels could be restored within 10 min. Recovery from an induced alkaline load was dependent on the chloride concentration in the external medium and sensitive to a stilbene derivative 4,4'-diisothiocyanatostilbene-2,2'-di-sulfonic acid that inhibits bicarbonate and chloride exchange. Therefore the recovery from alkalosis by hamster embryos appears to be via activity of the HCO(3)(-)/Cl(-) exchanger that was activated above a pH(i) set point of 7.24. In contrast, hamster oocytes and early 1-cell embryos (collected 3-4 h post-egg activation) could not recover from an intracellular alkalosis, and pH(i) remained elevated. Therefore, the hamster oocyte and the early 1-cell embryo still undergoing pronuclear formation lack an active HCO(3)(-)/Cl(-) exchanger for the restoration of pH(i). Inability to restore pH(i) from an alkali challenge resulted in a reduced ability of embryos to develop to the morula/blastocyst stages in culture, indicating that HCO(3)(-)/Cl(-) exchange is involved in physiological regulation of pH(i).