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We have interpreted the two-photon absorption spectrum of water-soluble copper indium sulfide (CIS) QDs with stoichiometry 0.18 (Cu), 0.42 (In), and 2 (S) and an average diameter of approximately 2.6 nm. For that, we employed the wavelength-tunable femtosecond Z-scan technique and the parabolic effective-mass approximation model, in which the excitonic transition energies were phenomenologically corrected due to the stoichiometry of the nanocrystal. This model considers a conduction band and three valence sub-bands allowing excitonic transitions via centrosymmetric (Δl = ±1, where l is the angular momentum of the absorbing state) and non-centrosymmetric (Δl = 0) channels. In such case, this became relevant because the CIS QDs with chalcopyrite crystalline structure is a non-centrosymmetric semiconductor. Thus, our experimental results pointed out two 2 PA allowed bands located at 715 nm (2hv = 3.47 eV) and 625 nm (2hv = 3.97 eV) with cross sections of (6.3 ± 1.0) x 102 GM and (4.5 ± 0.7) x 102 GM, respectively. According to the theoretical model, these 2 PA bands can be ascribed to the 1P1/2(h3) → 1S3/2(e) (lower energy band) and 1P1/2(hheavy) → 1S3/2(e) (90%)/(10%)1P1/2(hsplit-off) → 1P3/2(e) (higher energy band) excitonic transitions. A good agreement (magnitude and spectral position) between the experimental and theoretical data were obtained. However, our experimental data suggest that the higher-energy 2 PA band may have other contributions due to the mixing between the heavy- and the light-hole bands, which the effective mass model does not take into consideration.

Cyclosporine (CsA) blocks in vitro polyclonal activation of primary murine T-cells in a complex manner. This cannot be completely reversed by exogenous IL2, and leads to a partial blockade in expression of the IL2 receptor (p55 chain) and, more intensely, in CD69. In proliferation assays, T-cells recovered from CsA-treated cultures and washed free from CsA were markedly refractory to restimulation in the presence of fresh accessory cells. In cell titration restimulation assays, CsA-treated, but not control T-cells, were also markedly unresponsive to accessory cell-independent stimuli provided by immobilized anti-CD3 antibody or rIL2, combined to phorbol ester. CsA-treated, but not control activated T-cells, undergo progressive cell death after drug removal and reculturing. In contrast, primary T-cells activated by a CsA-resistant pathway (rIL2 plus phorbol ester) and treated with CsA, did not develop unresponsiveness, compared to controls. When primary T-cells were stimulated with rIL2 plus phorbol ester in the presence of the calcium ionophore ionomycin, treatment with CsA resulted in marked unresponsiveness of the T-cells, compared to untreated controls. The data indicate that primary activation of T-cells in vitro in the presence of CsA induces an unresponsive state which lasts independent of the presence of CsA, and results in progressive cell death. We suggest that these effects could characterize one additional mechanism of CsA action in vivo.

The roles of potassium and calcium in the slow hyperpolarizations of membranes of activated macrophages are investigated using standard intracellular electrical recording techniques. The amplitude of spontaneous slow hyperpolarizations decreases as a logarithmic function of the external potassium concentration in the culture medium. Similar dependence on the potassium gradient is observed when different levels of membrane potentials are imposed by constant current injection. The reversal potential for electrically evoked slow hyperpolarizations is −90 mV. A 10-fold increase in external potassium concentration causes a 60 mV shift of the reversal potential towards zero. Divalent cation ionophores (A23187 and X537A) can induce slow hyperpolarization responses in quiescent cells or permanent hyperpolarization in spontaneously active cells. The amplitude of the ionophore-induced hyperpolarizations is reduced by an increase in external potassium concentration in a manner consistent with data on slow hyperpolarization responses in the absence of ionophore. The calcium antagonist, verapamil, depresses the slow hyperpolarization responses at the concentration of 10−5 M. It is suggested that the development of the hyperpolarizing response is due to a calcium-dependent potassium channel. The data support the assumption that spontaneous and artificially elicited slow hyperpolarization responses share a common calcium-dependent mechanism.

Effects of four known blockers of T-lymphocyte potassium channels [verapamil, quinine, 4-aminopyridine (4-AP) and tetraethylammonium (TEA)], were studied on polyclonal T-cell activation induced by two plant mitogens (phytohemmaglutinin; PHA and concanavalin A; ConA), a mitogenic anti-Thy 1.2 monoclonal antibody (mAb G7) and phorbol ester (phorbol myristate acetate; PMA). Potassium channel blockers blocked all four modes of T-cell activation in a dose-dependent fashion with the same rank order of potency (verapamil greater than quinine greater than 4-AP greater than TEA). Kinetic studies of the timing of potassium channel blocker effect, indicated that, while 4-AP and TEA interfere only with early events of T-cell activation, verapamil and quinine can also interfere with later steps of T-cell mitogenesis. This notion was confirmed by studies of interleukin 2(IL-2)-directed activated T-cell growth. Verapamil and quinine blocked this late step in different types of activated T-cells with the same potency they blocked induction of resting T-cell mitogenesis. On the other hand, 4-AP and TEA, at maximal inhibitory doses for resting T-cells, showed little or no effect at IL-2-directed growth. Kinetic studies of the timing of quinine effect showed that the target of quinine action on activated T-cells is critically involved in IL-2 signalling within the first 2-4 h of IL-2 addition. These studies suggest that, besides the voltage-gated potassium channel previously described, a second target for verapamil and quinine action controls IL-2-derived signals to activated T-cells.

Electrophysiological properties of activated mouse macrophages cultured in vitro were studied using microelectrode techniques. In a high percentage of the individual cells analysed a slow hyperpolarization (SH) was observed with a concomitant decrease (2–4 times) of the input resistance. Increasing doses of tetraethyl ammonium progressively reduce the amplitude of the SH and at a concentration of 15 mM complete blockade of the phenomena is observed. Valinomycin, at a concentration of 10−7 M produces rapid and permanent hyperpolarization, with a shift in the membrane potential to about −50 mV. These data strongly support the previously proposed hypothesis that the development of SH is due to an increase in the membrane permeability to potassium ions.