Studies on predicting soil moisture levels at Andhra Loyola College, India, using SARIMA and LSTM models.

Time series modeling is a way to predict future values by examining temporal data. The present study analyzes the monthly mean soil moisture data at various depths: surface, profile, and root soil moisture, spanning from 1981 to 2022. The analysis employs two distinct approaches: the statistical seasonal autoregressive integrated moving average (SARIMA) and a deep learning long short-term memory (LSTM). The models are trained on a data set, covering the period from 1981 to 2021, acquired from the agricultural site at Andhra Loyola College in Vijayawada, Andhra Pradesh, India. Subsequently, the data from 2021 to 2022 is reserved for testing purposes. The study provides comprehensive insights into the design of both SARIMA and LSTM models, along with an evaluation of their performance using established error metrics such as the model mean absolute percentage error (MAPE), mean absolute error (MAE), and root mean squared error (RMSE). In the context of surface soil moisture prediction, the LSTM model demonstrates superior performance compared to SARIMA. Specifically, LSTM achieves a notably lower MAPE of 0.0615 in contrast to SARIMA's 0.1541, a reduced MAE of 0.0316 compared to 0.0871, and a diminished RMSE of 0.0412 as opposed to 0.1021. This pattern of enhanced accuracy persists across profile and root soil moisture predictions, further establishing LSTM's supremacy in predictive capability across diverse soil moisture levels.

Optimizing the luminescence efficiency of an europium (Eu3+) doped SrY2O4 phosphor for flexible display and lighting applications.

This research paper reports the synthesis and luminescence study of an Eu3+ activated SrY2O4 phosphor prepared by a modified solid-state reaction method with varying concentrations of Eu3+ ions (0.1-2.5 mol%). X-ray diffraction (XRD) revealed the orthorhombic structure and Fourier transform infrared spectroscopy (FTIR) methods were used to analyse the produced phosphors. Photoluminescence emission and excitation spectra were recorded for varying concentrations of Eu3+ ions, and an optimum concentration of 2.0 mol% was found to produce the highest intensity. Under 254 nm excitation the emission peaks were found to be at 580 nm, 590 nm, 611 nm and 619 nm, corresponding to transitions at 5D0 → 7F0, 5D0 → 7F1, and 5D0 → 7F2 respectively. Because of Eu3+ inherent luminosity, these emission peaks indicate radiative transitions between excited states of ions, making them useful for developing white light-emitting phosphors for optoelectronic and flexible display applications. The 1931 CIE (x, y) chromaticity coordinates were calculated from the photoluminescence emission spectra and found to be near white light emission, indicating the potential application of the prepared phosphor for light emitting diodes (white component). TL glow curve analysis was also performed for various concentrations of doping ions and UV exposure times, and a single broad peak was observed at 187 °C. Using the computerised glow curve deconvolution (CGCD) method, kinetic parameters were computed.

Fabrication of InVO4/SnWO4 heterostructured photocatalyst for efficient photocatalytic degradation of tetracycline under visible light.

In the present study, novel InVO4/SnWO4 nanocomposites with different concentrations of SnWO4 were successfully prepared using a facile hydrothermal technique and investigated employing a wide range of analytical methods for efficient photocatalytic degradation of tetracycline (TC). X-ray diffraction analysis showed the presence of the orthorhombic phases of both InVO4 and SnWO4 in the composite catalyst. Dispersion of SnWO4 nanoplates over the InVO4 nanosheets enhanced the synergistic interactions, improving the separation of charge carriers and their transfer. Furthermore, the formation of heterostructure expanded the absorption range and promoted visible light harvesting. The TC degradation efficiency of InVO4/SnWO4 nanocomposite (5 mg loading of SnWO4) reached 97.13% in 80 min under visible light, with the kinetic rate constants 5.51 and 7.63 times greater than those of pure InVO4 and SnWO4, respectively. Additionally, the scavenger results proved that hydroxyl radicals and holes played a significant role in the photodegradation of TC. Furthermore, the electrochemical impedance spectroscopy (EIS) and transient photocurrent response analysis showed enhanced e-/h+ partition efficiency. Thus, the formation of heterostructure with strong synergistic interactions can effectively transfer the excited charge carriers and shorten the reunion rate. Accordingly, the InVO4/SnWO4 nanocomposites exhibited remarkable photocatalytic performance due to the increased number of charge carriers on the surface.

Novel BiVO4-nanosheet-supported MoS2-nanoflake-heterostructure with synergistic enhanced photocatalytic removal of tetracycline under visible light irradiation.

This paper describes a simple in-situ hydrothermal technique for the production of BiVO4/MoS2 binary nanocomposites as visible-light-driven catalysts. The as-prepared samples were analyzed by structural, morphological, compositional, optical, surface area, and photocurrent analyses. The lattice fringe spaces at 0.304 nm and 0.612 nm were indexed to the (112) and (002) crystal planes of BiVO4 and MoS2, respectively. Antibacterial photocatalytic capabilities were assessed using tetracycline (TC). Consequently, it was observed that the BiVO4/MoS2 nanocomposite demonstrated improved antibacterial removal ability compared with the pristine samples. The BiVO4/MoS2 nanocomposite exhibited 97.46% removal of TC compared with the pure BiVO4 (43.76%) and MoS2 (35.28%) samples within 90 min. Thus, the photocatalytic performance was observed to follow the given order: BiVO4/MoS2 nanocomposite > BiVO4 > MoS2. The removal of TC after 90 min of irradiation was approximately 97.46%, 96.62%, 95.59%, and 94.45% after the 1st, 2nd, 3rd, and 4th cycles, respectively. Thus, the recycling tests revealed the stability of the photocatalyst, which exhibited a TC removal efficiency of 94.45% without distinct decay, even after the 4th cycle. According to the trapping results, hydroxyl radicals and holes were the key species and demonstrated a greater influence on the photocatalytic performance than superoxide radicals. The increased activity of the BiVO4/MoS2 nanocomposite may be attributed to its large surface area and tunable bandgap, which accelerate the charge-transport characteristics of the photocatalytic system. This insight and synergetic effects can provide a new approach for the development of novel heterostructure photocatalysts.

Luminescence studies of a Li2 Ca1-x SiO4 :xSm3+ phosphor for the generation of white light under NUV-excited phosphor converting LEDs.

In this paper, we present new aspects of Sm3+ -doped pure Li2 CaSiO4 as a suitable candidate for white light emitting diode (WLED) applications. The samples were mainly prepared using a conventional modified solid-state synthesis technique. The structural studies were done using X-ray diffraction and Rietveld refinement. Instruments such as a scanning electron microscope (SEM) were used to obtain information about the morphology of the as-prepared samples. Photoluminescence (PL) analysis of phosphor samples for variable concentrations of doping ions with variable excitations were presented. When doped with Sm3+ in host Li2 CaSiO4 it emitted intense blue, green and red emissions and a more intense red emission peak (605 nm) under 408 nm excitation (near-UV-blue). Our study shows that the as-prepared phosphor may be useful for optical devices and mainly for WLEDs. The corresponding transitions of doping ions and concentration quenching effect were studied in detail. The 1931 Commission Internationale de l'Eclairage (x, y) chromaticity coordinates showed the distribution of spectral regions calculated from PL emission spectra and this was found (0.63, 0.36) in the red region, so the phosphor may be useful for near-UV-blue excited WLED applications.

An effective CuO/Bi2WO6 heterostructured photocatalyst: Analyzing a charge-transfer mechanism for the enhanced visible-light-driven photocatalytic degradation of tetracycline and organic pollutants.

Over the past few years, industrial pollution has had a negative impact on aquatic life by releasing significant amounts of hazardous chemicals into the ecosystem. Therefore, it is imperative to develop photocatalytic materials with good photocatalytic activity and easy separation. Photocatalytic degradation has been employed for the removal of such contaminants using binary hybrid nanocomposites as photocatalysts. In the present study, binary CuO/Bi2WO6 (CuBW) nanocomposites with different loadings of Bi2WO6 (~5, 10, and 15 mg) were successfully constructed using a simple hydrothermal method and used as a potential photocatalyst for the degradation of tetracycline (TC) and methylene blue (MB) under visible-light irradiation. The structure, surface morphology, and optical properties were studied to investigate the formation of the heterostructure. Among the prepared samples, the CuBW nanocomposite containing the optimum content of Bi2WO6 (~10 mg) exhibited superior activity toward the photocatalytic degradation of TC (97.72%) in 75 min and MB (99.43%) in 45 min under visible-light illumination. Radical trapping experiments suggested that holes and •OH radicals were the dominant reactive species during the photocatalytic process. The photoelectrochemical results also confirmed the improved separation and transfer of electron-hole pairs at the interface of Bi2WO6 and CuO. Our results demonstrate that the binary CuO/Bi2WO6 nanocomposite has significant potential applications in the field of photocatalysis due to its enhanced separation of the photoexcited charge carriers and strong synergistic interactions.

Recent progress in transition metal oxide/sulfide quantum dots-based nanocomposites for the removal of toxic organic pollutants.

Water is an essential solvent that is extremely necessary for the survival of life. Water pollution due to the increased utilization of water for various processes, including domestic and industrial activities, poses a special threat that contaminates both surface and ground water. In recent years, advanced oxidation processes (AOPs) have been applied to deal with wastewater problems, which is a green method used to oxidize organic contaminants with strong oxidative radical species. Among the AOPs, photocatalytic technology is one of the most promising strategies for wastewater cleaning, which fulfills the aims of environmentally friendly and sustainable development. Owing to their unique electronic, optical, and structural properties, nanoscale semiconductors have received substantial interest as materials for AOPs, particularly inspired by their superb quantum confinement effects and large surface-area-to-volume ratio, which are essential for catalytic reaction kinetics. Recent advancements have revealed that semiconductor nanocrystals, known as quantum dots (QDs), are newly emerging zero-dimensional (0-D) nanomaterials, which have garnered much attention owing to their special physiochemical characteristics such as high conductivity, thermo-chemical and opto-mechanical stability, high adsorption coefficients, and, most importantly, their admirable recyclability. In this review, we provide a clear understanding of the importance of semiconductor QD-based nanocomposites in the degradation of organic pollutants, in addition to the mechanism involved in the reaction process. Following this, the enhancement of different materials, such as metal oxides and metal sulfide QD-based nanocomposites, is discussed in the context of combating environmental pollution.

ZnO nanosheets-decorated Bi2WO6 nanolayers as efficient photocatalysts for the removal of toxic environmental pollutants and photoelectrochemical solar water oxidation.

Herein we report the fabrication of novel Bi2WO6/ZnO heterostructured hybrids for organic contaminant degradation from wastewater and photoelectrochemical (PEC) water splitting upon solar illumination. The Bi2WO6/ZnO photocatalysts were synthesized using a simple and eco-friendly hydrothermal process without the support of any surfactants. From the photocatalytic experiments, heterostructured Bi2WO6/ZnO nanohybrid catalysts exhibited considerably better photocatalytic performance for rhodamine B (RhB) degradation under solar illumination. The BWZ-20 nanocomposite demonstrated superior photodegradation of RhB dye up to 99% in about 50 min. Furthermore, BWZ-20 photoelectrode showeda lower charge-transfer resistance than other samples prepared, suggesting its suitability for PEC water splitting. The photocurrent densities of Bi2WO6/ZnO photoelectrodes were evaluated under the solar irradiation. The BWZ-20 photoelectrode exhibited a significant photocurrent density (0.45 × 10-3A/cm2) at +0.3 V vs. Ag/AgCl, which was~1036-times higher than that of pure Bi2WO6, and ~4.8-times greater than the pure ZnO. Such improved photocatalytic and PEC activities are mainly attributed to the formation of an interface between ZnO and Bi2WO6, superior light absorption ability, low charge-transfer resistance, remarkable production of charge carriers, easy migration of charges, and suppression of the recombination of photogenerated charge carriers.

Effect of Al2O3 on structural and dielectric properties of PVP-CH3COONa based solid polymer electrolyte films for energy storage devices.

Nanocomposite polymer (NCP) films were prepared by doping sodium acetate (CH3COONa) in polymer of polyvinyl pyrrolidone (PVP) by complete dispersion of aluminum oxide (Al2O3) with different wt% proportions using solution cast method. The acquired NCP films were systematically characterized. The crystalline structure of the prepared NCP films was confirmed by XRD. The little agglomeration and grain sizes involved in the films were analyzed by SEM. The chemical bond formation and interchange reaction between the host, dopant salt and the nanofiller were confirmed by FTIR and Raman. The lowest energy bandgap values were observed to be 3.0 eV for the synthesized film with wt% ratio of PVP + CH3COONa:Al2O3 (80:20:1%). The highest ionic conductivity was found to be 1.05 × 10-3 S/cm for the prepared film with wt% ratio of PVP + CH3COONa:Al2O3 (80:20:1%). From the charge discharge characteristics it was concluded that the film with wt% ratio of PVP + CH3COONa:Al2O3 (80:20:1%) possesses long durability when compared to the other prepared films.

Spectroscopic investigations of Fe3+ doped poly vinyl alcohol (PVA) capped ZnSe nanoparticles.

Fe(3+) doped poly vinyl alcohol (PVA) capped ZnSe nanoparticles have been synthesized by using chemical method at room temperature. The prepared sample is characterized by X-ray diffraction, optical, photoluminescence (PL), electron paramagnetic resonance (EPR) and FT-IR techniques. Different physical parameters are evaluated by using measured values of refractive index and density. From the X-ray diffraction pattern, the average crystallite size is calculated by using Scherrer's formula and it is about 9 nm. Optical absorption spectrum reveals that the Fe(3+) ion enter into the lattice as octahedral symmetry. Crystal field (Dq) and inter-electronic repulsion parameters (B, C) are evaluated for Fe(3+) doped ZnSe nanoparticles as Dq=720, B=720 and C=2500 cm(-1). Photoluminescence spectrum of Fe(3+) doped ZnSe exhibits emission bands in UV and yellow regions. EPR spectrum showed various resonance signals at g=7.3, 3.5 and 2.0. FT-IR spectrum of Fe(3+) doped PVA capped ZnSe indicates the presence of O-H, C-H, C=C and C=O molecular groups.

Knee stabilisation following infected knee arthroplasty with bone loss and extensor mechanism impairment using a modular cemented nail.

Infected Total Knee Replacement with significant bone loss and loss of extensor mechanism poses a difficult management problem. Arthrodesis relying on bony union can be difficult to achieve and can result in significant limb shortening. We retrospectively looked at the outcome of seven patients with significant bone loss and extensor mechanism insufficiency following infected TKR who underwent knee stabilisation using a modular cemented nail. The nail relied on the strong coupling mechanism between the modular femoral and tibial components. Pain score improved from a mean of 7.9 pre-operatively to 1.5 post-operatively at a mean follow up of 39.6 months (range 7-68) months. Two patients underwent technically easy revision nailing for recurrent infection and aseptic loosening. The Endo-Model(R) Knee Fusion Nail (Newsplint, UK/Waldemar Link, GmbH & Co. KG, Hamburg, Germany) has good early results in terms of pain relief and provides a stable knee in cases with significant bone loss and extensor mechanism insufficiency following an infected TKR thus avoiding an above knee amputation.

Cost and efficiency of HIV voluntary counselling and testing centres in Andhra Pradesh, India.

BACKGROUND: [corrected] As part of the effort to control HIV/AIDS, the number of HlV voluntarycounselling and testingcentres (VCTCs) is increasing rapidly in the public health system of the Indian state of Andhra Pradesh, which is estimated to have one of the highest rates of HIV infection in India. However, systematic data on the cost and efficiency of providing VCT services in India are not available to help guide efficient use of resources for these services. METHODS: We used standardized methods to obtain detailed cost and output data for the 2002-03 fiscal year from written records and interviews in 17 VCTCs in the public health system in Andhra Pradesh. We calculated the economic cost per client receiving VCT services, and analysed the variation and determinants of total and unit costs across VCTCs. We used multivariate regression techniques to estimate incremental unit costs. We assessed hurdles towards serving an optimal number of clients by VCTCs. RESULTS: In the 2002-03 fiscal year, 32 413 clients received the complete sequence of services at the 17 VCTCs, including post-HIV test counselling. The number of clients served by each VCTC ranged from 334 to 7802 (median 979). The overall HIV-positive rate in post-test counselled clients was 20.5% (range 5.4%-52.6%). The cost per client for the complete VCT sequence varied 6-fold between VCTCs (range Rs 141.5-829.6 [US 2.92-17.14 dollars], median Rs 363.5 [US 7.51 dollars]). The cost per client was significantly lower at VCTCs with more clients (p < 0.001, R2 = 0.83; power function) due to substantial fixed costs. Personnel made up the largest component of cost (53.7%). The cost per client had a significant direct relation with percent personnel cost for VCTCs (p < 0.001, R2 = 0.58; exponential function). A multiple regression model revealed that the incremental cost of providing complete VCT services to each HIV-positive and -negative client was Rs 123.5 (US 2.54 dollars) and Rs 59.2 (US 1.22 dollars), respectively. Fourteen VCTCs (82.4%) reported that they could serve more clients with the available personnel and infrastructure, and that inadequate demand for their services was the main hurdle towards achieving this. CONCLUSION: These data suggest that the efforts of the National AIDS Control Organisation of India and the Andhra Pradesh State AIDS Control Society in increasing VCTCs could yield even higher benefit if the demand for these services was enhanced, as this would increase the number of clients served and reduce the cost per client. Ongoing systematic cost-efficiency analysis is necessary to help guide efficient use of HIV-control resources in India.

Role of abrasion of the femoral component in revision knee arthroplasty.

We carried out 60 revision procedures for failed porous coated anatomic total knee replacements in 54 patients, which were divided into two groups. The 14 knees in group I had a well-fixed femoral component at surgery which was retained, and in the 46 knees in group II both tibial and femoral components were loose and were revised using a variety of implants. Our review comprised clinical and radiological assessment. A total of 13 knees required a second revision. Six (42%) in group I failed very early (mean 2.1 years) when compared with seven (15%) in group II (mean 6.8 years). Failure was due to wear of the polyethylene insert by the abraded, retained femoral component (crude odds ratio 4.07; 95% CI 1.07 to 15.5). We recommend a complete change of primary bearing surfaces at the time of revision of an uncemented total knee replacement in order to prevent early wear of polyethylene.

Can breast surgeons read mammograms of symptomatic patients in the one-stop breast clinic?

AIM: To establish how accurate surgeons were when compared to the radiologists in interpreting symptomatic mammograms in one-stop clinics. METHODS: The surgeons were asked to write their opinion on the mammograms which was compared with the radiologists' report. 144 patients were involved in the study and the data were analysed by McNemara's test for paired categorical data. RESULTS AND CONCLUSIONS: Surgeons were accurate in interpreting most of the mammographic findings. However, they underestimated the presence of benign calcification which was statistically significant. Surgeons can, therefore, be involved in double reading of mammograms in symptomatic breast disease patients and improve the sensitivity which has been the case in double reading by radiologists in the breast screening programme.

Differences in Ca(2+) signaling underlie age-specific effects of secretagogues on colonic Cl(-) transport.

Taurodeoxycholic acid (TDC) stimulates Cl(-) transport in adult (AD), but not weanling (WN) and newborn (NB), rabbit colonic epithelial cells (colonocytes). The present study demonstrates that stimuli like neurotensin (NT) are also age specific and identifies the age-dependent signaling step. Bile acid actions are segment and bile acid specific. Thus although TDC and taurochenodeoxycholate stimulate Cl(-) transport in AD distal but not proximal colon, taurocholate has no effect in either segment. TDC increases intracellular Ca(2+) concentration ([Ca(2+)](i)) in AD, but not in WN and NB, colonocytes. In AD cells, TDC (5 min) action on Cl(-) transport needs intra- but not extracellular Ca(2+). NT, histamine, and bethanechol increase Cl(-) transport and [Ca(2+)](i) in AD, but not WN, distal colonocytes. However, A-23187 increased [Ca(2+)](i) and Cl(-) transport in all age groups, suggesting that Ca(2+)-sensitive Cl(-) transport is present from birth. Study of the proximal steps in Ca(2+) signaling revealed that NT, but not TDC, activates a GTP-binding protein, Galpha(q), in AD and WN cells. In addition, although WN and AD colonocytes had similar levels of phosphatidylinositol 4,5-bisphosphate, NT and TDC increased 1,4,5-inositol trisphosphate content only in AD cells. Nonresponsiveness of WN cells to Ca(2+)-dependent stimuli, therefore, is due to the absence of measurable phospholipase C activity. Thus delays in Ca(2+) signaling afford a crucial protective mechanism to meet the changing demands of the developing colon.

Janus kinase 2 (JAK2) regulates prolactin-mediated chloride transport in mouse mammary epithelial cells through tyrosine phosphorylation of Na+-K+-2Cl- cotransporter.

Epithelial chloride (Cl-) transport is achieved by the coordinated action of symporters such as the Na+-K+-2Cl- cotransporter (NKCC1) and chloride channels such as the cystic fibrosis transmembrane conductance regulator (CFTR). As a secretory tissue, mammary epithelial cells are obvious candidates for such mechanisms, but Cl- transport and its hormonal regulation have been poorly delineated in mammary epithelial cells. We determined whether the mammary epithelial cell line, HC11, transports chloride and whether this was regulated by PRL, a hormone known to stimulate ion transport. HC11 cells express both CFTR and NKCC1. Exposure to PRL or PGE1 increased Cl- transport in HC11 cells. This was inhibited by the NKCC1 blocker, furosemide, and by the Cl- channel inhibitor, diphenylamine 2-carboxylate. Dose and time course of PRL action indicate that PRL had maximal effect on Cl- transport at 1 microg/ml and at 10 min of stimulation. Examination of the signaling pathways suggests that the PRL effect on Cl- transport does not involve an increase in [Ca2+]i or MAP kinase activity. RT-PCR analyses indicate that HC11 cells express mRNA for Janus kinase 1 (JAK1), JAK2, and signal transducer and activator of transcription 5 (STAT5) but not for JAK3. PRL treatment of HC11 cells increased phosphorylation of STAT5. The JAK2 inhibitor AG490 blocked phosphorylation of STAT5 and PRL-induced, but not PGE1-induced, Cl- transport. NKCC1, but not CFTR, is tyrosine phosphorylated in HC11 cells. PRL enhanced tyrosine phosphorylation of NKCC1, and this effect was attenuated by the JAK2 inhibitor AG490. These results are the first demonstrations of a role for tyrosine phosphorylation of NKCC1 and of the PRL-JAK2 cascade in the regulation of Cl- transport.

Evidence for the presence of cGMP-dependent protein kinase-II in human distal colon and in T84, the colonic cell line.

Heat-stable enterotoxin (STa) stimulates intestinal Cl(-) secretion by activating guanylate cyclase C (GCC) to increase intracellular cyclic GMP (cGMP). In the colon, cGMP action could involve protein kinase (PK) G-II or PKA pathways, depending on the segment and species. In the human colon, both PKG and PKA pathways have been implicated, and, therefore, the present study examined the mechanism of cGMP-mediated Cl(-) transport in primary cultures of human distal colonocytes and in T84, the colonic cell line. Both cell preparations express mRNA for CFTR, Na(+)-K(+)-2Cl(-) cotransporter (NKCC1), GCC and PKG-II as detected by RT-PCR. The effects of STa and the PKG-specific cGMP analogues, 8Br-cGMP and 8pCPT-cGMP, on Cl(-) transport were measured using a halide-sensitive probe. In primary human colonocytes and T84 cells, STa, the cGMP analogues and the cAMP-dependent secretagogue, prostaglandin E(1) (PGE(1)), enhanced Cl(-) transport. The effects of 8Br-cGMP and 8pCPT-cGMP suggested the involvement of PKG, and this was explored further in T84 cells. The effects of 8pCPT-cGMP were dose-dependent and sensitive to the PKG inhibitor, H8 (70 microM), but H8 had no effect on PGE(1)-induced Cl(-) secretion. In contrast, a PKA inhibitor, H7 (50 microM), blocked PGE(1)-mediated but not 8pCPT-cGMP-induced Cl(-) transport. 8pCPT-cGMP enhanced phosphorylation of the PKG-specific substrate, 2A3, by T84 membranes in vitro. This phosphorylation was inhibited by H8. These results strongly suggest that cGMP activates Cl(-) transport through a PKG-II pathway in primary cells and in the T84 cell line of the human colon.

Ion transport during growth and differentiation.

The major function of the adult colon is to reabsorb fluid from the chyme. This ability to conserve salt and water is especially important in newborns, where reserves are small and diarrhea is frequent. Although much is known about regulation of Cl- transport in the adult colon, postnatal changes in electrolyte transport are not well characterized. We have established an in vitro model to study colonic epithelial cells (colonocytes) at different stages of development. Primary cultures were isolated from newborn, weanling, and adult rabbit colon and properties such as growth and Cl- transport characterized. The isolation procedure yielded a crypt-enriched population of cells, and the cell yield per gram mucosa increased with age. The colonocytes also showed an age-related decrease in attachment to extracellular matrix, with maximum attachment seen with Matrigel and collagen IV. The crypt enrichment was confirmed by demonstrating that the cell population was capable of transporting Cl-, which was stimulated by agents such as forskolin and phorbol esters at all ages. Agents that increased intracellular cGMP, however, did not increase Cl- transport at any age. It was interesting to observe that the secondary bile acid, taurodeoxycholate, stimulated Cl- transport only in the adult but not newborn or weanling distal colonocytes. We have demonstrated that rabbit distal colonocytes can be kept viable in culture and transport Cl- at all ages. However, the regulation of Cl- transport changes during ontogeny and depends on the signaling pathway.

Regulation of intestinal epithelial brush border Na(+)/H(+) exchanger isoforms, NHE2 and NHE3, in C2bbe cells.

Until recently, studies to characterize the intestinal epithelial Na(+)/H(+) exchangers had to be done in nonepithelial, mutated fibroblasts. In these cells, detection of any Na(+)/H(+) exchange activity requires prior acid loading. Furthermore, most of these experiments used intracellular pH changes to measure NHE activity. Because changes in pH(i) only approximate Na(+)/H(+) exchange activity, and may be confounded by alterations in buffering capacity and/or non-NHE contributions to pH regulation, we have used (22)[Na] unidirectional apical to cell uptake to measure activities specific to NHE2 or NHE3. Furthermore, we performed these measurements under basal, nonacid-stimulated conditions to avoid bias from this nonphysiological experimental precondition. Both brush border NHEs, when expressed in the well-differentiated, intestinal villuslike Caco-2 subclone, C2bbe (C2), localize to the C2 apical domain and are regulated by second messengers in the same way they are regulated in vivo. Increases in intracellular calcium and cAMP inhibit both isoforms, while phorbol ester affects only NHE3. NHE2 inhibition by cAMP and Ca(++) involves changes to both K(Na) and V(max). In contrast, the same two second messengers inhibit NHE3 by a decrease in V(max) exclusively. Phorbol ester activation of protein kinase C alters both V(max) and K(Na) of NHE3, suggesting a multilevel regulatory mechanism. We conclude that NHE2 and NHE3, in epithelial cells, are basally active and are differentially regulated by signal transduction pathways.

A unique Na+/H+ exchanger, analogous to NHE1, in the chicken embryonic fibroblast.

We report the characterization of an Na+/H+ exchanger (NHE) in embryonic fibroblasts (SL-29 cells) of the chicken, a terrestrial vertebrate, where Na+ conservation is important. This exchanger is electroneutral, has a single Na+ binding site, and is highly sensitive to amiloride (IC50 2 microM), dimethyl amiloride (350 nM), and ethyl-isopropyl amiloride (25 nM). It is stimulated by serum, transforming growth factor-alpha, hypertonicity, and okadaic acid. Although these features make it resemble mammalian NHE1, other characteristics suggest distinct differences. First, in contrast to mammalian NHE1 it is inhibited by cAMP and shows a biphasic response to phorbol esters and a highly variable response to increased intracellular Ca2+ concentration. Second, whereas full-length human and rat NHE1 cDNA probes recognize a 4.8-kb transcript in rat tissues, they recognize only a 3.9-kb transcript in chicken tissues. An antibody against amino acids 631-746 of human NHE1 sequence fails to recognize a protein in SL-29 cells. Rat NHE2 and NHE3 probes do not recognize any transcript in chicken fibroblasts. The SL-29 exchanger differs markedly from the previously characterized chicken intestinal apical exchanger in its amiloride sensitivity and regulation by phorbol esters. These results suggest that a modified version of mammalian NHE1 is present in chicken tissues and imply that another functionally distinct Na+/H+ exchanger is expressed in aves.