Modulation of calcium-activated chloride current via pH-induced changes of calcium channel properties in cone photoreceptors.

The activity of calcium-activated chloride channels is controlled through the complex interaction of cellular mechanisms affecting calcium entry, buffering, and extrusion, and an unknown stoichiometric relation between intracellular Ca concentration and Cl channel activation. Here, we show that calcium-activated chloride current [ICl(Ca)] in cone photoreceptors is also highly sensitive to external pH, being strongly reduced by acidification and enhanced by alkylinization of the bathing medium. We propose that this modulation is accounted for by the pH sensitivity of Ca channel activation and permeation, already well characterized in other cells, which we now extend to cone photoreceptor Ca channels. Acidification of the external medium from a control pH of 7.4 shifts the Ca channel activation range positively by about 10 mV at pH 6.8, reducing the magnitude of calcium current with a consequent reduction of chloride current. Alkylinization shifts the Ca channel activation range negatively by about 8 mV at pH 8 and produces larger calcium currents during step depolarizations that in turn elicit larger chloride tail currents. Modulation of ICl(Ca) by pH suggests other consequences of the pH-induced shift in Ca channel gating, for one, modification of Ca-dependent transmitter release, which could be especially significant in photoreceptors where the cell's operating voltage range overlaps only the lower end of the Ca channel activation range.     

Modulation of high-dose methotrexate toxicity by a non-toxic level of 5-fluorouracil

High-dose methotrexate (MTX) toxicity is reduced by a non-toxic dose of 5-fluorouracil (FU) when these agents are used in combination. Changes in the hematopoietic system (platelets, erythrocytes, leukocytes, hemoglobin, and hematocrit), ileal tissue, body weight, and mean survival were used as parameters to assess toxicity. For all parameters studied, there were no significant differences between the scheduling of MTX (245 mg/kg) after a priming dose of FU (25 mg/kg), simultaneous MTX and FU, FU alone, and control. However, sequential treatment with MTX followed by FU, and MTX alone resulted in: a marked decrease in the hematopoietic parameters; significant morphological changes in ileal tissue; a reduction of body weight; and increased mortality of animals. Hence, this study suggests that FU, a cytotoxic agent, may protect against MTX toxicity and improve its therapeutic index when FU administration precedes MTX or when these agents are given simultaneously.     

无家可归者多地点结核病传播

地点：2002-2003年，在华盛顿King县，结核病（TB）在多个收容所的人群中大规模爆发。 目的：控制结核病在多个地点的传播。设计：2002年，对收容所病人的接触者进行筛查，作结核菌素皮试（TSTs）和症状回顾。根据这些筛查的结果，确定和优选传播的地点。2003年，对暴露于这些点的队列作彻底的筛查（例如，症状回顾，TST，胸部X线检查[CXR]，痰检和痰培养）。利用PCR为基础的方法对从病人那里分离出来的结核分枝杆菌作基因分型，以快速确认爆发相关病人。 结果：2002-2003年，King县313例确诊的病人中有48例（15％）与爆发相关；通过基因分型，47例培阳病人分离出和爆发相匹配的菌株。3个收容所由于在2002年接纳的病人超过12人，所以人群中TST阳性率（约30％）高于收容所中的一般水平（7％）。用一个痰培养筛查接触者和CXR发现结核病人的敏感度相似（分别为77％和62％）。 结论：一个广泛的资源密集的途径可能有助于控制疾病的传播。这次爆发突显出无家可归者的脆弱性和在城市维持强有力的结核病规划的必要性。     

Genetic diversity among clinical isolates of Acremonium strictum determined during an investigation of a fatal mycosis

Primarily saprophytic in nature, fungi of the genus Acremonium are a well-documented cause of mycetoma and other focal diseases. More recently, a number of Acremonium spp. have been implicated in invasive infections in the setting of severe immunosuppression. During the course of routine microbiological studies involving a case of fatal mycosis in a nonmyeloablative hematopoietic stem cell transplant patient, we identified a greater-than-expected variation among strains previously identified as Acremonium strictum by clinical microbiologists. Using DNA sequence analysis of the ribosomal DNA intergenic transcribed spacer (ITS) regions and the D1-D2 variable domain of the 28S ribosomal DNA gene (28S), the case isolate and four other clinical isolates phenotypically identified as A. strictum were found to have <99% homology to the A. strictum type strain, CBS 346.70, at the ITS and 28S loci, while a sixth isolate phenotypically identified only as Acremonium sp. had >99% homology to the type strain at both loci. These results suggest that five out of the six clinical isolates belong to species other than A. strictum or that the A. strictum taxon is genetically diverse. Based upon these sequence data, the clinical isolates were placed into three genogroups.     

CD1d deficiency exacerbates inflammatory dermatitis in MRL-lpr/lpr mice

Mechanisms responsible for the development of autoimmune skin disease in humans and animal models with lupus remain poorly understood. In this study, we have investigated the role of CD1d, an antigen-presenting molecule known to activate natural killer T cells, in the development of inflammatory dermatitis in lupus-susceptible MRL-lpr/lpr mice. In particular, we have established MRL-lpr/lpr mice carrying a germ-line deletion of the CD1d genes. We demonstrate that CD1d-deficient MRL-lpr/lpr mice, as compared with wild-type littermates, have more frequent and more severe skin disease, with increased local infiltration with mast cells, lymphocytes and dendritic cells, including Langerhans cells. CD1d-deficient MRL-lpr/lpr mice had increased prevalence of CD4(+) T cells in the spleen and liver and of TCR alpha beta (+)B220(+) cells in lymph nodes. Furthermore, CD1d deficiency was associated with decreased T cell production of type 2 cytokines and increased or unchanged type 1 cytokines. These findings indicate a regulatory role of CD1d in inflammatory dermatitis. Understanding the mechanisms by which CD1d deficiency results in splenic T cell expansion and cytokine alterations, with increased dermal infiltration of dendritic cells and lymphocytes in MRL-lpr/lpr mice, will have implications for the pathogenesis of inflammatory skin diseases.     

Detection of penicillin-resistant Streptococcus pneumoniae with commercially available broth microdilution panels.

We compared penicillin MICs obtained with three different commercially available broth microdilution panels (MicroScan, Sensititre, and Pasco) with MICs obtained with reference microdilution panels for 20 well-characterized pneumococci with decreased susceptibilities to penicillin (7 resistant and 13 intermediate). All panels were supplemented with 2 to 5% lysed horse blood (LHB) prepared in-house. Additional supplements included fastidious inoculum broth (FIB) for MicroScan panels and commercially prepared LHB (Difco) for Pasco panels. The percentages of penicillin-resistant strains (MIC 2 micrograms/ml) detected by the different methods follow: MicroScan-FIB, 0; MicroScan-LHB 0; Pasco in-house LHB, 71; and Sensititre-LHB, 100. The percentages of intermediate strains (MIC = 0.1 to 1.0 micrograms/ml) detected by the different methods follow: MicroScan-FIB, 31; MicroScan-LHB 23; Pasco in-house LHB, 46; and Sensititre-LHB, 85. Difco LHB supplement failed to support the growth of 86% of the strains in the Pasco panels. Of the commercially available panels evaluated, only Sensititre, supplemented with LHB prepared in-house could reliably detect penicillin-resistant pneumococci.