Studies in African Cyperaceae 21. New taxa and combinations in Kyllinga Rottb.

Eight new species, one new subspecies and one new variety of Kyllinga are described from tropical Africa, viz. Kyllinga brunneoalba Lye, K. afropumila Lye, K, albogracilis Lye, K. aureovillosa Lye, K. alba–purpurea Lye, K. rhizomafragilis Lye, K. microbracleata Lye, K. afro–occidentalis Lye, K. erecta Schumacher ssp. albescens Lye, and K. melanosperma Nees var. hexalata Lye. In addition nine new combinations are made, viz. Kyllinga peteri (Kükenthal) Lye, K. bigibbosa (Fosberg) Lye, K. odorata Vahl ssp. appendiculata (K. Schum.) Lye, K. alba Nees ssp. ascolepidioides (H. Cherm.) Lye, K. nervosa Steudel ssp.flava (C.B.C1.) Lye, K. melanosperma Nees ssp. elata (Steudel) Lye, K. brevifolia Rottb. ssp. lurida (Kükenthal) Lye, K. bre–vifolia Rottb. ssp. intricata (H. Cherm.) Lye, and K. comosipes (Mattf. & Kükenthal) Agnew & Hanid ssp. decolorans (Kükenthal) Lye.     

Specificity of tetraethylammonium and quinine for three K channels in insulin-secreting cells

Summary The effects of tetraethylammonium (TEA) and quinine on Ca-activated [K(Ca)]. ATP-sensitive [K(ATP)]K channels and delayed-rectifier K current [K(dr)] have been studied in cultured insulin-secreting HIT cells using the patch-clamp technique. K(Ca) and K(ATP) channels were identified in excised, outside/ out patches using physiological solutions and had unitary conductances of 60.8±1.3 pS (n=31) and 15.4±0.3 pS (n=40). respectively. Macroscopic K(dr) current (peak current=607±100 pA at +50 mV,n=14) were recorded in the presence of 100 m cadmium and 0.5 m tetrodotoxin. Tetraethylammonium (TEA) blocked all three channel types but was more effective on K(Ca) channels (EC₅₀=0.15mm) than on K(ATP) channels (EC₅₀=15mm) or K(dr) currents (EC₅₀=3mm). Quinine also blocked all three currents but was less effective on K(Ca) channels (EC₅₀=0.3mm) while equally effective against K(ATP) channels and K(dr) currents (EC₅₀=0.025mm). TEA blocked K(Ca) and K(ATP)_channels by reducing their single-channel conductances and decreasing the probability of K(ATP) channel opening. Quinine blocked K(Ca) channels by reducing the single-channel conductance, but blocked K(ATP) channels by reducing the probability of channel opening. Reinterpretation of previous microelectrode studies in light of these findings suggest that, (i) only K(ATP) channels are active in low glucose, (ii) both K(Ca) and K(dr) channels may assist Ca-spike repolarization, and (iii) K(Ca) channels play no role in forming the burst pattern of Ca spiking in the B cell.     

Coordination fluxionality of copper(II) with a sterically constrained Pyrazole-containing chelating ligand. crystal and molecular structure at 298 K and 140 K of=1,6-Bis(3,5-dimethylpyrazol-1-yl)-2,5-dimethyl-2,5-diazahexane-bis(thiocyanato-N)copper(II), [C18H28CuN8S2]

The X-ray structure determinations of the compound Cu(debd)(NCS)₂ at 140 K and at 298 K showed at both temperatures a tetragonal crystal system with space group P4₁2₁2 (Z=4) and a molecular C₂ symmetry. Cell dimensions are: a= 9.577, c=24.422 Å and V=2240 ų at 140 K, and a=9.614, c=24.854 Å and V=2297 ų at 298 K. 1218 (140 K) and 1057 (298 K) reflections with I > 1.96 σ(I) were used for the solution to a final R_w value of 0.036 (140 K) and 0.032 (298 K). The copper(II) environment is roughly octahedral. The copper to nitrogen bonds vary with temperature: Cu-N(amine)=2.242 Å (140 K) and 2.191 Å (298 K), Cu-N(pyrazole)=2.099 Å (140 K) and 2.193 Å (298 K), and Cu-NCS=2.060 Å (140 K) and 2.034 Å (298 K). The ESR spectra of the Cu(II)- doped isomorphous Zn(II) and Cd(II) compounds, which have been measured at ambient temperature, at 77 K, and at 3 K, clearly reflect the temperature dependence of the structure.     

Substrate kringle-mediated catalysis by the streptokinase-plasmin activator complex: Critical contribution of kringle-4 revealed by the mutagenesis approaches

Streptokinase (SK) is a protein co-factor with a potent capability for human plasminogen (HPG) activation. Our previous studies [1] have indicated a major role of long-range protein-protein contacts between the three domains (alpha, beta, and gamma) of SK and the multi-domain HPG substrate (K1-K5CD). To further explore this phenomenon, we prepared truncated derivatives of HPG with progressive removal of kringle domains, like K5CD, K4K5CD, K3-K5CD (K3K4K5CD), K2-K5CD (K2K3K4K5CD) and K1-K5CD (K1K2K3K4K5CD). While urokinase (uPA) cleaved the scissile peptide in the isolated catalytic domain (μPG) with nearly the same rate as with full-length HPG, SK-plasmin showed only 1-2% activity, revealing mutually distinct mechanisms of HPG catalysis between the eukaryotic and prokaryotic activators. Remarkably, with SK.HPN (plasmin), the ‘addition’ of both kringles 4 and 5 onto the catalytic domain showed catalytic rates comparable to full length HPG, thus identifying the dependency of the “long-range” enzyme-substrate interactions onto these two CD-proximal domains. Further, chimeric variants of K5CD were generated by swapping the kringle domains of HPG with those of uPA and TPA (tissue plasminogen activator), separately. Surprisingly, although native-like catalytic turnover rates were retained when either K1, K2 or K4 of HPG was substituted at the K5 position in K5CD, these were invariably lost once substituted with the evolutionarily more distant TPA- and uPA-derived kringles. The present results unveil a novel mechanism of SK.HPN action in which augmented catalysis occurs through enzyme-substrate interactions centered on regions in substrate HPG (kringles 4 and 5) that are spatially distant from the scissile peptide bond.     

The effects of drug complexation on the stability and conformation of human serum albumin

We report different analytical methods used to study the effects of 3\'-azido-3\'-deoxythymidine, aspirin, taxol, cisplatin, atrazine, 2,4-dichlorophenoxyacetic, biogenic polyamines, chlorophyll, chlorophyllin, poly(ethylene glycol), vanadyl cation, vanadate anion, cobalt-hexamine cation, and As2O3, on the stability and secondary structure of human serum albumin (HSA) in aqueous solution, using capillary electrophoresis, Fourier transform infrared, ultraviolet visible, and circular dichroism (CD) spectroscopic methods. The concentrations of HSA used were 4% to 2% or 0.6 to 0.3 mM, while different ligand concentrations were 1 microM to 1 mM. Structural data showed drugs are mostly located along the polypeptide chains with both specific and nonspecific interactions. The stability of drug-protein complexes were in the order K(VO(2+)) 1.2 x 10(8) M(-1) > K(AZT) 1.9 x 10(6) M(-)1 > K(PEG) 4.1 x 10(5) M(-1) > K(atrazine) 3.5 x 10(4) M(-1) > K(chlorophyll) 2.9 x 10(4) M(-1) > K2,4-D 2.5 x 10(4) M-1 > K(spermine) 1.7 x 10(4) M(-1) > K(taxol) 1.43 x 10(4) M(-1) > K(Co(3+)) > 1.1 x 10(4) M(-1) > K(aspirin) 1.04 x 10(4)i(-1) > K(chlorophyllin) 7.0 x 10(3) M(-1) > K(VO(3)(-)) 6.0 x 103 M(-1) > K(spermidine) 5.4 x 10(3) M(-1) > K(putrescine) 3.9 x 10(3) M(-1) > K(As(2)O(3)) 2.2 x 10(3) M(-1)> K(cisplatin) 1.2 x 10(2) M(-1). The protein conformation was altered (infrared and CD results) with major reduction of alpha-helix from 60 to 55% (free HSA) to 49 to 40% and increase of beta-structure from 22 to 15% (free HSA) to 33 to 23% in the drug-protein complexes. The alterations of protein secondary structure are attributed to a partial unfolding of HSA on drug complexation.     

Measurements in potassium-supplemented athletes during and after hypokinetic and ambulatory conditions

Hypokinesia (diminished movement) induces significant potassium (K) changes; however, little is known about K deposition and deficiency during hypokinesia (HK). Using K supplements during and after HK, the aim was to establish body K deposition and K deficiency during HK. Studies were done during the pre-HK period of 30 d, HK period of 364 d, and post-HK period of 30 d. Forty male trained athletes aged 24.9 ± 8.0 y were chosen as subjects. They were equally divided into four groups: unsupplemented active control subjects (UACS), unsupplemented hypokinetic subjects (UHKS), supplemented active control subjects (SACS), and supplemented hypokinetic subjects (SHKS). Hypokinetic subjects were limited to an average walking distance of 0.7 km/d. Control subjects ran an average distance of 11.6 km/d. The SHKS and SACS groups took 95.0 mg elemental K/kg body weight daily. Fecal K excretion, urinary sodium (Na) and K excretion, plasma K and Na levels, plasma renin activity (PRA), plasma aldosterone (PA), food and fluid intake, and physical characteristics were measured. During HK, fecal K loss, urinary K and Na loss, and plasma K, Na, PRA, and PA levels increased significantly (p ≤ 0.05), whereas during the initial days of post-HK, the levels of the measured parameters decreased significantly (p ≤ 0.05) in the SHKS and UHKS groups as compared with the SACS and UACS groups, respectively. During HK, body weight, body fat, peak oxygen uptake, food and fluid intake decreased significantly (p ≤ 0.05), whereas during the initial days of post-HK period remained significantly (p ≤ 0.05) depressed and fluid intake increased in SHKS and UHKS groups when compared with the SACS and UACS groups, respectively. However, during HK and post-HK plasma, urinary, and fecal K changed significantly (p ≤ 0.05) more in the SHKS group than in the UHKS group. The deposition of K was significantly (p ≤ 0.05) lower and K deficiency much higher in the SHKS group than in the UHKS group. Fecal K loss, urinary K and Na loss, plasma K, Na, PRA, and PA levels, body weight, body fat, peak oxygen uptake, and food and fluid intake did not change significantly in the SACS and UACS when compared with their baseline control values. It was shown that plasma K concentration and urinary and fecal K excretion increased during HK and decreased significantly (p ≤ 0.05) during post-HK. post-HK. Oral K supplements did not influence plasma or fecal and urinary K either during HK or post-HK. It was concluded that the low plasma K level and fecal and urinary K loss during post-HK may indicate the presence of K deficiency, and increased K in plasma, urine, and feces during HK and in the presence of K deficiency may suggest the body’s inability to retain K during HK.     

Simultaneous Congo red decolorization and electricity generation in air-cathode single-chamber microbial fuel cell with different microfiltration, ultrafiltration and proton exchange membranes

Different microfiltration membrane (MFM), proton exchange membrane (PEM) and ultrafiltration membranes (UFMs) with different molecular cutoff weights of 1 K (UFM-1K), 5 K (UFM-5K) and 10 K (UFM-10K) were incorporated into air-cathode single-chamber microbial fuel cells (MFCs) which were explored for simultaneous azo dye decolorization and electricity generation to investigate the effect of membrane on the performance of the MFC. Batch test results showed that the MFC with an UFM-1K produced the highest power density of 324 mW/m² coupled with an enhanced coulombic efficiency compared to MFM. The MFC with UMF-10K achieved the fastest decolorization rate (4.77 mg/L h), followed by MFM (3.61 mg/L h), UFM-5K (2.38 mg/L h), UFM-1K (2.02 mg/L h) and PEM (1.72 mg/L h). These results demonstrated the possibility of using various membranes in the system described here, and showed that UFM-1K was the best one based on the consideration of both cost and performance.     

Different maternal genome donor to Kengyilia species inferred from chloroplast trnL-F sequences

To reveal the maternal donor of species in genus Kengyilia, the chloroplast trnL-F sequences of 14 Kengyilia species and several related diploid species were analyzed by using Maximum Parsimony (MP) and Bayesian Inference (BI) methods. The species in Kengyilia were clustered in different clades, which indicated that Agropyron (P) is the likely maternal genome donor to Kengyilia melanthera, K. mutica and K. thoroldiana, while the maternal donor to Kengyilia batalinii, K. nana, K. kokonorica, K. kaschgarica, K. hirsuta, K. alatavica, K. gobicola, K. zhaosuensis, K. rigidula, K. longiglumis and K. grandiglumis was St or Y Roegneria genome.     

Breeding behaviour of Kunzea pomifera (Myrtaceae): self-incompatibility, intraspecific and interspecific cross-compatibility

To examine breeding system characteristics of the endemic Australian prostrate shrub Kunzea pomifera, artificial hybridisations were undertaken using thirteen different genotypes of K. pomifera, to elucidate: (1) self-incompatibility, (2) intraspecific cross-compatibility in the species and (3) interspecific cross-compatibility with each of K. ambigua and K. ericoides. K. pomifera exhibited very low self-compatibility, with the barrier to self-fertilisation being prevention of pollen-tube growth in the style or ovary. Following intraspecific pollination amongst a number of different genotypes of K. pomifera, 38.4% of pollinated flowers developed fruit; arrest of compatible pollen-tubes in the style, preventing fertilisation, contributes to the low fruit set in this species. Interspecific compatibility was examined between K. pomifera (pistillate parent) and K. ambigua (staminate parent) where seed set per pollinated flower (4.47) was not significantly different from intraspecific crosses (4.66). In crosses between K. pomifera (pistillate parent) and K. ericoides as staminate plant, 0.037% of pollinated flowers produced fruit, with 0.0075 seeds per pollinated flower. Reproductive barriers between these two species were evident in the style of K. pomifera, where the growing tips of the K. ericoides pollen-tubes swelled and ceased to grow.     

Importance of product/reactant equilibration in the kinetics of the phosphoglucose isomerization reaction by differential stopped flow microcalorimetry

The kinetics for the isomerization of fructose-6-phosphate to glucose-6-phosphate (F6P --> G6P) by baker's yeast phosphoglucose isomerase (PGI) with regard to k(cat) and K(m) were determined from analysis of differential stopped flow microcalorimeter measurements using the integrated form of the Michaelis-Menten rate equation. Values for K(m) (F6P --> G6P) that were determined at pH 8.0 and ionic strength 0.1M at 293.4, 298.4, 303.4, and 311.5K exhibited a linear dependence on the substrate concentration at each temperature because of the substrate-product equilibrium. The minimum values for K(m) ranged from 2.62+/-0.55 mM at 293.4K to 7.8+/-4.8mM at 311.5K and were the same as the minimum values for the reverse reaction (G6P --> F6P) at 293.4 K and 298.4 K. Minimum values for k(cat) increased with temperature, from 2.78+/-0.34s(-1) at 293.4K to 11.4+/-1.0s(-1) at 311.5K, and for the reverse reaction, G6P --> F6P, from 0.852+/-0.086 s(-1) at 293.4K to 1.46+/-0.06s(-1) at 298.4K. The enzyme efficiency at 311.5K is close to the collision rate for a diffusion-controlled process in solution. The [F6P]/[G6P] equilibrium constants were determined from comparison of the values of k(cat) in both directions and were 0.307+/-0.053 at 293.4K and 0.395+/-0.033 at 298.4K. The heats of reaction in the F6P --> G6P direction increased from -8.96+/-0.26 kJmol(-1) at 311.5K to -8.27+/-0.40 kJmol(-1) at 293.4K, a value in fair agreement with 7.01+/-0.32 kJmol(-1) in the opposite G6P --> F6P direction.     

Potassium nutrition of rice (Oryza sativa L.) varieties under NaCl salinity

A salt-tolerant (Pokkali) and a salt-sensitive (IR28) variety of rice (Oryza sativa L.) were grown in a phytotron to investigate the effect of K (0, 25, 50 and 75 mg K kg^–1 soil) application on their salt tolerance. Potassium application significantly increased potential photosynthetic activity (Rfd value), percentage of filled spikelets, yield and K concentration in straw. At the same time, it also significantly reduced Na and Mg concentrations and consequently improved the K/Na, K/Mg and K/Ca ratios. IR28 responded better to K application than Pokkali. Split application of K failed to exert any beneficial effect over basal application.     

The application of high-performance liquid chromatography for the resolution of proteins encoded by the human adenovirus type 2 cell transformation region

The human adenovirus 2 (Ad2) transformation genes are located in early region E1a (map position (mp) 1.3–4.5) and E1b (mp 4.6–11.2) on the linear duplex Ad2 DNA genome of M_r 23 × 10⁶ (viral DNA is divided into 100 map units). E1b codes for three major proteins of apparent molecular weights 53,000 (53K), 19K, and 20K; smaller quantities of 21K, 22K, and 23K proteins that are related to 53K are also synthesized in Ad2-infected cells. Because the resolution and purification of these Ad2 candidate transformation proteins proved very difficult by conventional protein purification methods, the applicability of high-performance liquid chromatography (HPLC) methodology was examined. Starting with a crude cytoplasmic S100 fraction of Ad2-infected human cells, the resolution of the Ad2 E1b-coded 19K, 20K, 21K, 22K, and 23K proteins by reverse-phase HPLC using a C₈ column and a linear 0–60% 1-propanol gradient in 0.5 m pyridine formate was achieved, E1b proteins purified under these conditions retained their immunological reactivity. By anion-exchange HPLC using a linear 10 mm to 1 m NaCl gradient in 10 mm 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid buffer, pH 7.6, the same five Ad2 E1b-coded 19K–23K proteins were separated, with improved resolution of the 19K protein. Based on these findings, protocols for the extensive purification of the E1b-19K and E1b-20K proteins have been developed. These results illustrate the potential of HPLC methodology for the rapid purification of biologically interesting proteins from complex cellular mixtures of proteins.     

Potassium uptake efficiency and dynamics in the rhizosphere of maize (Zea mays L.), wheat (Triticum aestivum L.), and sugar beet (Beta vulgaris L.) evaluated with a mechanistic model

Plant species differ in nutrient uptake efficiency. With a pot experiment, we evaluated potassium (K) uptake efficiency of maize (Zea mays L.), wheat (Triticum aestivum L.), and sugar beet (Beta vulgaris L.) grown on a low-K soil. Sugar beet and wheat maintained higher shoot K concentrations, indicating higher K uptake efficiency. Wheat acquired more K because of a greater root length to shoot dry weight ratio. Sugar beet accumulated more shoot K as a result of a 3- to 4-fold higher K influx as compared to wheat and maize, respectively. Nutrient uptake model NST 3.0 closely predicted K influx when 250 mg K kg^?1 were added to the soil, but under-predicted K influx under low K supply. Sensitivity analysis showed that increasing soil solution K concentration (C_Li) by a factor of 1.6–3.5 or buffer power (b) 10- to 50-fold resulted in 100% prediction of K influx. When both maximum influx (I_max) and b were increased by a factor of 2.5 in maize and wheat and 25 in sugar beet, the model could predict measured K influx 100%. In general, the parameter changes affected mostly calculated K influx of root hairs, demonstrating their possible important role in plant K efficiency.     

Differential regulation of keratin 8 and 18 messenger RNAs in differentiating F9 cells

F9 embryonal carcinoma cells (F9EC) can be induced to differentiate in vitro into epithelial cells expressing keratin 8 (K8) and keratin 18 (K18). cDNAs corresponding to K8 and K18 mRNAs were cloned and used to study the change in the abundance of these mRNAs during differentiation of F9 cells into parietal endoderm-like cells by treatment with retinoic acid (RA) or with RA and dibutyryl cAMP (Bt₂cAMP). Using an RNase protection assay, it was determined that K8 mRNA was induced slightly before K18 mRNA and that it accumulated to a greater extent than K18 mRNA. Furthermore, differentiation in presence of Bt₂cAMP plus RA resulted in an earlier induction of the two mRNAs and a higher level of expression of K8 mRNA. These results indicate that K8 and K18 mRNAs are regulated differently in F9 cells.     

异育银鲫幼鱼对饲料中维生素K需求的研究

以不同维生素K水平(0.13、2.15、3.25、6.40、12、17.20和23.20 mg/kg饲料)的7种精制饲料喂养初始体重约为(2.17±0.01) g的异育银鲫(Carassius auratus gibelio)10周, 每个处理3个重复, 研究异育银鲫对维生素K的需求量。结果显示: 饲料中维生素K的添加可以明显降低摄食率, 饲料中维生素K含量为2.15 mg/kg时, 摄食率出现最大值, 之后显著下降(P<0.05), 在12 mg/kg时达到最低值。特定生长率随着维生素K的添加表现出升高的趋势, 饲料中维生素K含量为12 mg/kg时, 出现最大值, 但是差异不显著(P>0.05)。饲料中维生素K的含量从0.13 mg/kg升至3.25 mg/kg时, 饲料效率显著升高(P<0.05), 随着饲料中维生素K的进一步添加, 趋于稳定(P>0.05), 在12 mg/kg时达到最大值, 并且与特定生长率呈正相关关系(SGR=0.01 FE+0.95, R²=0.95)。血液红细胞数目随着饲料维生素K含量的增加先显著升高(P<0.05), 在6.40 mg/kg时达到最大值, 之后趋于稳定(P>0.05)。血红蛋白含量、血球容积比、血清钙含量与血液中红细胞数目表现出相似的趋势, 均在不添加维生素K组出现最低值, 但是差异不显著(P>0.05)。肝体比、肥满度及鱼体生化组成均不受饲料维生素K水平的影响(P>0.05)。分别对饲料效率、红细胞数目进行折线回归得出异育银鲫幼鱼对维生素K的最适需求量为3.73—6.72 mg/kg饲料。     

Biosystematic studies of Kengyilia melanthera and K. kokonorica (Poaceae: Triticeae)

Kengyilia melanthera (Keng)J. L. Yang, Yen et Baum and K. kokonorica (Keng)J. L. Yang, Yen et Baum are two hexaploid perennial grasses of the tribe Triticeae native in west China. K. melanthera and K. kokonorica were hybridized with Roegneria kamoji Ohwi(2n=42,StStHHYY) and K. hirsuta (Keng)J. L. Yang, Yen et Baum (2n = 42, PPStStYY) respectively. Chromosome pairing behaviour at metaphase I in the parents and hybrids was studied. Meiotic configurations were 18.20 Ⅰ + 11.74 Ⅱ + 0.09 Ⅲ + 0.01V for R. kamoji×K. melanthera, 1.06Ⅰ + 20.47 Ⅱ for K. hirsuta×K. melanthera, 19.36Ⅰ + 11.26 Ⅱ + 0.04Ⅲ for R. kamoji×K. kokonorica, and2.46Ⅰ + 19.44Ⅱ + 0.14 Ⅲ + 0.06 Ⅳ for K. hirsuta×K. kokonorica. Considering chromosome pairing in the hybrids, as well as morphological characters, K. melanthera and K. kokonorica should be grouped in Kengyilia Yen et J. L. Yang instead of being keeped inRoegneria sect. Paragropyron Keng , or in Elymus L. or Elytrigia Desv.     

Rice yield in relation to electroultrafiltration extractable soil potassium

Summary Potassium in wetland rice soils from five different locations in the Philippines was analyzed using the electroultrafiltration (EUF) technique and by extraction withN NH₄ acetate (pH 7). The soils contained low exchangeable K and responded to K application. The K soil test values were calibrated against the rice response to K application under field conditions. EUF extractable soil K correlated highly significantly with the rice yield response to K fertilizer, whereas the NH₄ acetate extractable K (exchangeable K) did not. Under limiting K supply in soils, rice yield depends more on the EUF-K than on the exchangeable K. Maximum grain yields were obtained when the EUF-K values after harvest and before wetland preparation were above 30 ppm K.     

Kinetic comparison of ouabain-resistant K:CI fluxes (K:Cl [Co]-transport) stimulated in sheep erythrocytes by membrane thiol oxidation and alkylation

Summary The stimulatory effects of two thiol (SH) group oxidants, methylmethane thiosulfonate (MMTS) and diazene dicarboxylic acid bis [N,N-dimethylamide] (diamide), on the kinetics of ouabain-resistant (OR) K:Cl [co]-transport in low K (LK) sheep red blood cells were compared with the effects of alkylating agents, notably N-ethylmaleimide (NEM). At low concentrations, both MMTS and diamide stimulated K:CI [co]-transportv and with a latency period, as measured by OR zero-trans K efflux and OR uptake of external Rb, Rb_o, as K congener in Cl and NO₃ media. At high concentrations the effect of diamide saturated, and that of MMTS disappeared. The stimulatory effect of MMTS was partially reversed by the reducing agent dithiothreitol (DTT) known to fully restore the diamide-activated K flux (Lauf, J. Memb. Biol. 101:179–188, 1988). In diamide pre-equilibrated LK sheep red cells, the K_m of K:Cl [co]-transport for external Cl, Cl_o, was 84.3 mM, and 18.7 mM for Rb_o, with nearly identical V_max values around 4 mmol Rb/L cells × h for K (Rb) fluxes in Cl and after correction for the small Cl-independent component. Zero net K (Rb) flux existed at K_c (cell K)/Rb_o concentration ratios, [K]_c/[Rb]_c, of 0.8 i.e. when the electrochemical driving forces across the membrane were about equal. The measured K efflux/Rb influx ratios were almost twice those predicted from [K]_c/[Rb]_o and the Cl equilibrium potential suggesting that the diamide-stimulated K (Rb) flux may occur through non-diffusional, carrier-mediated transport. The effects of NEM and of A23187 plus/minus Ca or chelators on K: [co]Cl-transport (Lauf, Am. J. Physiol. 249:C271–278, 1985) consisted primarily of V_max changes. Thus, all chemical interventions resulted in an increase of the number of actively transporting K:Cl [co]-transport units or an augmented turnover number per existing site.