Reduction of forkhead box P3 levels in CD4+CD25high T cells in patients with new-onset systemic lupus erythematosus

The aim of this study was to quantify and evaluate the forkhead box P3 (FoxP3) expression regulatory T cells in new-onset systemic lupus erythematosus (SLE) patients before and after treatment. Forty-four newly diagnosed and untreated SLE patients, including 24 with active disease (SLEDAI > or = 10) and 20 with inactive disease (SLEDAI < 5), were enrolled in this study. Twenty-one age- and sex-matched healthy volunteers were also included as controls. Peripheral blood samples were collected and mononuclear cells isolated. The expression of CD25 and FoxP3 in CD4(+) T cells were analysed with flow cytometry. CD4(+)CD25(+) (3.95-13.04%) and CD4(+)CD25(high) (0.04-1.34%) T cells in peripheral blood in untreated patients with new-onset active lupus were significantly lower than that in the patients with inactive lupus (7.27-24.48%, P < 0.05 and 0.14-3.07% P < 0.01 respectively) and that in healthy controls (5.84-14.84%, P < 0.05). Interestingly, the decrease in CD4(+)CD25(high) T cells was restored significantly in patients with active lupus after corticosteroid treatment. There was, however, a significantly higher percentage of CD4(+)FoxP3(+) T cells in patients with active (5.30-23.00%) and inactive (7.46-17.38%) new-onset lupus patients compared with healthy control subjects (2.51-12.94%) (P < 0.01). Intriguingly, CD25 expression in CD4(+)FoxP3(+) T cells in patients with active lupus (25.24-62.47%) was significantly lower than that in those patients with inactive lupus (30.35-75.25%, P < 0.05) and healthy controls (54.83-86.38%, P < 0.01). Most strikingly, the levels of FoxP3 expression determined by mean fluorescence intensity in CD4(+)CD25(high) cells in patients with active SLE were significantly down-regulated compared with healthy subjects (130 +/- 22 versus 162 +/- 21, P = 0.012). CD4(+)CD25(high) T cells are low in new-onset patients with active SLE and restored after treatment. Despite that the percentage of CD4(+)FoxP3(+) T cells appear high, the levels of FoxP3 expression in CD4(+)CD25(high) T cells are down-regulated in untreated lupus patients. There is a disproportional expression between CD25(high) and FoxP3(+) in new-onset patients with active SLE.     

Topography of 3H-DHA, 3H-QNB, 3H-Dopamine, and 3H-DAGO Binding Sites Distribution in the Central Part of the Sinoatrial Node in Rat Heart

The topography of distribution of ³H-dihydroalprenolol, ³H-quinucledinyl benzilate, ³H-dopamine, and ³H-DAGO binding sites in the central part of the sinoatrial node in rat heart was studied by autoradiography after electrophysiological identification of the dominant pacemaker region location. Receptor asymmetry between the lateral and median regions of the central part of the sinoatrial node was shown. The dominant pacemaker region lay in the lateral area of the sinoatrial node; the number of binding sites for all four ligands was minimum in it. The number of binding sites gradually increased in the cranial and caudal directions from the dominant pacemaker region along the sinoatrial node artery (more smoothly in the caudal direction). The relative densities of bindings sites for ³H-dihydroalprenolol and ³H-dopamine were higher in the lateral region compared to the perinodal working myocardium, while the densities for ³H-quinucledinyl benzilate and ³H-DAGO were virtually the same. The distribution of binding sites along the artery in the median region of the sinoatrial node was even for ³H-quinucledinyl benzilate and ³H-DAGO. For ³H-DAGO these parameters were close to those in the perinodal atrial myocardium, for ³H-quinucledinyl benzilate somewhat lower. Curves presenting the distribution of binding site densities for ³H-dihydroalprenolol and ³H-dopamine in the median region of the sinoatrial node were similar, with a pronounced peak in the region contralateral to the dominant pacemaker region, and significantly higher binding parameters compared to those for the perinodal atrial myocardium. The difference consisted in higher density of ³H-dopamine binding sites in the median region of the sinoatrial node in comparison with the lateral region. Binding activity was maximum in the wall of the sinoatrial node artery. The distribution of binding sites for ligands to the main autonomic nervous system neurotransmitters in the rat heart sinoatrial node is heterogeneous. __________ Translated from Byulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 140, No. 10, pp. 472–477, October, 2005     

Electron microscopic radioautographic study on the syntheses of DNA, RNA and protein in the livers of aging mice

The syntheses of DNA, RNA and protein in the livers of aging mice was studied by electron microscopic radioautography using radioactive precursors. Labeling with³H-thymidine was observed in the nuclei of some hepatocytes at various prenatal and postnatal ages. The percentage of labeled cells decreased after birth, then slowly fell to the lowest value at 2 years. The silver grains with³H-uridine labeling were observed in both the nuclei and cytoplasm of hepatocytes at various ages. There was a peak in uridine labeling at 14 days, and then it slowly decreased until old age. The number of silver grains with³H-leucine labeling in the cytoplasm of hepatocytes was small. It increased after birth, reached the maximum at 1 month, and continued to decrease with aging.     

Galectin-1 reduction and changes in T regulatory cells may play crucial roles in patients with unexplained recurrent spontaneous abortion

To investigate the changes of Galectin-1 and T-lymphocyte phenotypes in unexplained recurrent spontaneous abortion (URSA). Totally 60 participants were recruited and divided into 3 groups in average: pregnant patients with URSA (URSA group), normal early pregnant women with induced abortion (IA group) and normal non-pregnant women (control group). After the tissue and blood sample were collected, Galectin-1 was measured using enzyme-linked immunosorbent assay. Then the proportion of T regulatory cells was determined by flow cytometry. The expression levels of Galectin-1 in IA group and URSA group was significantly higher than that in the control group (24.30 ± 3.06 and 6.23 ± 2.41 vs. 1.30 ± 0.66, P < 0.05). Besides, the expression level of Galectin-1 in URSA group was lower than that in IA group (P < 0.05). The percentage of CD4⁺CD25⁺Foxp3⁺ Tregs was lower in URSA group than IA group (0.77 ± 0.31 vs. 1.00 ± 0.35, P < 0.05) and the ratio of CD4⁺CD25⁺Foxp3⁺/CD4⁺ in URSA group was also obviously lower than that in IA and control group (P < 0.05). Galectin-1 and CD4⁺CD25⁺Foxp3⁺ may play essential roles in maintaining a normal pregnancy and their reduction may involve in the pathogenesis of URSA.     

Intratumoral FOXP3VEGFR2 regulatory T cells are predictive markers for recurrence and survival in patients with colorectal cancer

Previously, we have shown that CD8⁺T/FOXP3⁺ cell ratio but not FOXP3⁺ cell number alone is an independent prognostic factor for colorectal cancer. In the present study, we evaluated whether the number of intratumoral FOXP3⁺VEGFR2⁺ (itFOXP3⁺VEGFR2⁺) T cells alone could be a predictive factor for survival prognosis in patients with colorectal cancer. Distribution of regulatory T cells (Tregs) at tumor sites derived from 88 patients with primary colorectal cancer was fluorescence-immunohistochemically examined. Relatively low number of itFOXP3⁺VEGFR2⁺ cells significantly correlated with poor disease-free survival (DS) and overall survival (OS); multivariate analysis indicated that number of itFOXP3⁺VEGFR2⁺ cells is an independent predictive and prognostic factor of DS and OS while neither intratumoral FOXP3⁺ cell number nor intratumoral FOXP3⁺VEGFR2⁻ cell number alone showed significant correlation with DS or OS. These results suggest that FOXP3⁺VEGFR2⁺ may be a better predictive Treg marker than FOXP3⁺ alone for recurrence and survival in patients with colorectal cancer.     

Distribution of labeled amino acids and delta sleep-inducing peptide in the body after instillation into the rabbit conjunctiva

P. K. Anokhin Research Institute of Normal Physiology, Academy of Medical Sciences of the USSR. Institute of Molecular Genetics, Academy of Sciences of the USSR. I. M. Sechenov First Moscow Medical Institute. Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 110, No. 9, pp. 236–237, September, 1990.     

The reduced proportion of New splenic T-cells in the zinc-deficient growing rat is not due to increased susceptibility to apoptosis

Dietary zinc deficiency has been associated with an increased risk of infection. It has been reported that zinc-deficient rats have fewer New T-cells (TCRαβ⁺CD90⁺) compared to diet-restricted and control rats, which over time could adversely affect the ability of the organism to fight off infections. We hypothesized that the lower proportion of New T-cells in zinc deficiency is due to an increased susceptibility to apoptosis. Weanling, Sprague Dawley rats were assigned to one of four dietary treatment groups for 3 weeks: zinc-deficient (ZD, <1 mg zinc/kg, ad libitum), diet-restricted (DR, 30 mg zinc/kg, limited to the amount of feed as consumed by ZD), marginally zinc-deficient (MZD, 10 mg zinc/kg, ad libitum) or control (CTL, 30 mg zinc/kg, ad libitum). Thymocytes and splenocytes were labeled for flow cytometric determination of cell surface markers and DNA staining (for simultaneous determination of the phenotype of apoptotic cells) and assessed by Western blotting for apoptotic markers. Cells were analyzed immediately, or after incubation for 7 h with or without dexamethasone. There was no difference in the proportion of CD90⁺ thymocytes; however ZD rats had a higher proportion of Cytotoxic (CD90⁺4⁻8⁺) thymocytes compared to MZD and CTL. ZD had a lower proportion of splenic New T-cells compared to DR, MZD and CTL. There was no effect of diet on the proportion of apoptotic thymocytes or splenocytes, except ZD splenoctyes had a lower Bax/Bcl-xl ratio compared to DR and CTL. We characterized the splenic New T-cells into Helper and Cytotoxic subsets and found that ZD had a higher ratio of Helper to Cytotoxic New T-cells compared to MZD and CTL. These results do not support the hypothesis of increased apoptotic removal of New T-cells in ZD in growing rats. The regulation of CD90 expression should be explored in future studies.     

Die Auswirkung der sekundären Nebennierenrindeninsuffizienz auf Körperwasser und austauschbare Elektrolyte

Zusammenfassung Bei 5 Patienten mit bestätigter sckundärer Nebennierenrindeninsuffizienz wurden das Ganzkörperwasser, der Extracellulärraum, das austauschbare Natrium und das austauschbare Kalium mit Hilfe von³H₂O,⁷⁷Br,²⁴Na und⁴³K vor und nach Entzug der Nebennierenrinden-substitution (7,5 mg Prednison tägl.) bestimmt. Bei allen Patienten kam es zur Abwanderung von Wasser und Natrium aus dem Extracellulärraum in die Zellen. Da das austauschbare Natrium (und das austauschbare Kalium) unverändert blieben, läßt sich die bisherige Theorie, daß eine Abnahme des Körpernatriums für die beobachtete Verkleinerung des Extracellulärraumes verantwortlich sei, auf Grund unserer Resultate bei Patienten mit sekundärer Nebennierenrinden-insuffizienz nicht bestätigen.     

Electron microscopic radioautographic study on the nucleic acids and protein syntheses in the aging mouse testis

The ultrastructural difference between³H-thymidine labeled and unlabeled germ cells in mouse testis was observed by electron microscopic radio-autography. The RNA and protein syntheses in the seminiferous tubules of aging mice were also studied using³H-uridine and leucine radioautography. At embryonic day 19, the Sertoli cells and myoid cells labeled with³H-thymidine were often observed. The gonocytes labeled with³H-thymidine that were not rich in cell organelles appeared from the early postnatal stage. The silver grains were located mainly over the nuclei of labeled gonocytes, but a few were over the mitochondria. The labeling index of spermatogonia for³H-thymidine rapidly increased from 2 weeks on, and stayed relatively constant until old age. The labeling indices of the other two kinds of cells, i.e., Sertoli and myoid cells, decreased to low levels with aging, while these cells continued RNA and protein syntheses through aging, as detected by the incorporation of³H-leucine and uridine.     

Epidermal growth factor stimulates Ca2+ uptake of human erythrocytes

To examine the functional significance of epidermal growth factor (EGF) binding sites present on the human erythrocyte membrane [Engelmann et al. (1992) Am J Hematol 39:239–241], the effect of EGF on ⁴⁵Ca²⁺ uptake and on ²²Na⁺ efflux from these cells has been studied. In all cases media contained 1.25 mM Ca²⁺, whereas Na⁺ and K⁺ were varied. In 140 mM Na⁺/5 mM K⁺ medium EGF (250 ng/ml) stimulated ⁴⁵Ca²⁺ uptake by 50%–90% in quin-2-loaded cells, and by up to threefold in untreated cells. Increasing extracellular K⁺ up to 75 mM at the expense of extracellular Na²⁺ stimulated the EGF-induced ⁴⁵Ca²⁺ uptake by about twofold compared to 145 mM Na⁺ medium both in quin-2-loaded and in untreated cells. In 145 mM K⁺ medium, however, no EGF-induced ⁴⁵Ca²⁺ uptake was detectable in quin-2-loaded cells, while in untreated cells Ca²⁺ entry was stimulated twofold by EGF. After increasing intracellular Na⁺ from 6 mmol/l cells to 18 mmol/l cells in untreated cells suspended in 145 mM K⁺ medium, ⁴⁵Ca²⁺ uptake induced by EGF gradually increased. In contrast, in 140 mM Na⁺/5 mM K⁺ as well as in 70 mM Na⁺/75 mM K⁺ medium, ⁴⁵Ca²⁺ uptake accelerated by EGF was largely unaffected by a modified red cell Na⁺ content. When ²²Na-loaded untreated red cells were suspended in 145 mM K⁺ medium EGF stimulated red cell ²²Na⁺ efflux by more than threefold. In 140 mM Na⁺/5 mM K⁺ as well as in 70 mM Na⁺/75 mM K⁺ medium, no ²²Na⁺ efflux induced by the growth factor was evident. The results are consistent with the idea that EGF stimulates (at least) two components of ⁴⁵Ca²⁺ uptake in human erythrocytes. One of the two is unmasked in 145 mM K⁺ medium, inhibited by quin-2 loading, accelerated by intracellular Na⁺ and appears to involve reversed Na⁺/Ca²⁺ exchange.     

Influence of Mitragyna Ciliata (MYTA) on the Microsomal Activity of ATPase Na+/K+ Dependent Extract on a Rabbit Heart

Mitragyna ciliata (MYTA) (Rubiaceae) inhibits plasmodia activity. MYTA induces a cardiotonicity of the digitalic type on rat''s isolated heart. In this work we studied the effect of MYTA on microsomal Na⁺/K⁺ dependant ATPase (Na⁺, K⁺ ATPase) extracted from the heart of a rabbit since digitalics inhibit Na⁺, K⁺ ATPase. Our results revealed that the Na⁺/K⁺ ATPase has an optimum pH of 7.4 and temperature of 37°C respectively. There is a linear relationship between the organic phosphate formed and the incubation time over 25 mins incubation period. The ATP hydrolysis rate in the presence of MYTA was 0.775 µM/min. LINEWEAVER and BURK plots showed that MYTA did not alter K_M (1.31 mM) but decreased V_MAX. This study shows that MYTA exerts a non-competitive inhibition on the microsomal Na⁺/K⁺ ATPase extracted from rabbit heart with a Ci₅₀ of 48 µg / ml. We conclude that the mechanism of action of MYTA is linked to the inhibition of the Na⁺/K⁺ ATPase like cardiotonics of the digitalic type.     

Murine CD5+, CD8+ Normal Fetal Liver Cells Enhance an Immune Response: Benzo(α) Pyrene-Exposed CD5+ Fetal Liver Cells are Inhibitors

The effects of 150 ug benzo(α)pyrene/gm body weight given intraperitoneally to the pregnant mouse at mid-gestation leads to long-lasting (> 18 months post-birth) immune deficiency in the progeny. Although the progeny are immunodeficient their T cell subsets induced marked enhancement and/or inhibition of cell proliferation in an immune response. Earlier we saw a striking increase (up to 7-fold) in CD8⁺ cells in the 18 day gestation liver of benzo(α)pyrene-exposed fetuses. We hypothesized that immune deficiency in carcinogen-exposed progeny could be attributed to an increase in classical CD8⁺ suppressor T-cells. Surprisingly, however, we found that CD8⁺ and CD5⁺ (Lyt 1⁺) cells of normal fetal liver enhance cell proliferation in an immune response. However, liver CD5⁺ cells from benzo(α)pyrene-exposed fetuses led to a dramatic reduction of the enhancing effect. Thus, as a novel finding, it appears that the profile of CD5⁺ cells, under the ontogenic influence of benzo(α)pyrene, transforms from cells that normally augment cell proliferation in an immune response to cells that are inhibitors. On the other hand the functional status of CD8⁺ cells is not affected. This change in physiological status of CD5⁺ cells may have important implications on T and B cell interactions, and the role of CD5⁺cells in T cell receptor signaling.     

Effect of high NaCl intake on Na+ and K+ transport in the rabbit distal convoluted tubule

Morphological studies have demonstrated that a chronic increase in distal Na⁺ delivery causes hypertrophy of the distal convoluted tubule (DCT). To examine whether high NaCl-intake also causes functional changes in the well defined DCT, we measured transmural voltage (V _T), lumen-to-bath Na⁺ flux (J _Na(LB)), and net K⁺ secretion (J _K(net)) in DCTs obtained from control rabbits and those on high NaCl-intake diets. The lumen negativeV _T was significantly greater in the high NaCl group than in the control group. The net K⁺ secretion (pmol mm^–1 min^–1) was greater in the high NaCl-intake group (54.1±13.0 vs 14.7±5.6). The K⁺ permeabïlities in both luminal and basolateral DCT membranes, as assessed by the K⁺-induced transepithelial voltage deflection inhibitable with Ba²⁺, were increased in the experimental group. The lumen-to-bath²²Na flux (pmol mm^–1 min^–1) was also greater in the experimental group (726±119 vs 396±65). TheV _T component inhibitable with amiloride was also elevated in the high NaCl-intake group. Furthermore, Na⁺–K⁺-ATPase activity of the DCT was higher in the experimental than in the control group. We conclude that high NaCl intake increases both Na⁺ reabsorption and K⁺ secretion by the DCT. This phenomenon is associated with an increased Na⁺–K⁺-ATPase activity along with increased Na⁺ and K⁺ permeabilities of the luminal membrane, and an increase in the K⁺ permeability of the basolateral membrane. Cellular mechanisms underlying these functional changes remain to be established.     

Molecular origin of Na/Li exchanger: Evidence against the involvement of major cloned erythrocyte transporters

Numerous studies have demonstrated heightened Na⁺/Li⁺ countertransport (NLCT) activity in erythrocytes of patients with essential hypertension or diabetic nephropathy. The same carrier also contributes to the therapeutic action of lithium salt, widely used in the treatment of psychiatric disorders. However, the molecular origin of NLCT remains unknown. This study examined the role of major ion transporters in NLCT by comparative analysis of its activity and that of ion transporters providing inwardly directed ⁸⁶Rb, ²²Na and ³²P fluxes. NLCT was below the detection limit in rat erythrocytes and ∼50-fold higher in rabbits compared to humans. Unlike NLCT, the activities of Na⁺,K⁺-ATPase, Na⁺,K⁺,2Cl⁻ cotransporter and anion exchanger were somewhat similar in the erythrocytes of these species, whereas Na⁺,P_i cotransport was in 1:2:6 proportion in rats, humans and rabbits, respectively. Loading of erythrocytes with Li⁺ for NLCT measurement did not affect the activity of Na⁺,P_i cotransporter. Keeping in mind that NLCT is much higher in rabbits vs humans and rats, we compared the set of membrane proteins in these species using 2-dimensional gel electrophoresis. This approach revealed 174 common spots, whereas 132 proteins were detected only in human and rabbit erythrocyte membranes. Among these proteins, we found 17 spots whose expression was higher by more than 5-fold in rabbit compared to human erythrocytes. Thus, our results argue against the involvement of major ion transporters in NLCT. They also show that comparative proteomics is a potent tool to identify the molecular origin of this carrier.     

A kinetic analysis of the facilitatory action of adrenaline

The²²Na⁺-efflux from skeletal muscle cells of frog (Rana nigromaculata) was measured in Ringer solutions containing different concentrations of K⁺ (0.1 to 30 mM). The effects of adrenaline (30 M) and ouabain (10 M) on the²²Na⁺-efflux were investigated for the purpose to clarify the mechanism of the facilitatory effect of adrenaline on Na⁺–K⁺ pump. The rate coefficient for the ouabain-sensitive²²Na⁺-efflux increases with increasing extracellular K⁺ concentrations and adrenaline potently facilitates these rate coefficients. On the basis of Michaelis-Menten type kinetics assumed for the reaction between pump site and extracellular K⁺, it is concluded that adrenaline decreases the dissociation constant (K_m), and increases the maximum Na⁺-efflux.This work was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan.     

Platelet survival determined with51Cr Versus111In

Summary Optimal labelling conditions of human platelets with¹¹¹In-oxine were determined in vitro. Based on this optimized technique, platelet mean life span (MLS) and platelet sequestration site were comparatively evaluated in 79 patients with two labels,⁵¹Cr (n=26) and¹¹¹In (n=53). Patients were subgrouped according to clinical criteria as autoimmune thrombocytopenic purpura (AITP) (group 1;n=49), hypersplenism (2;n=12), impaired thrombopoiesis (3;n=3), unclassified thrombocytopenia (4;n=6), and nonthrombocytopenic patients (5;n=9). In patients with AITP and hypersplenism the mean values for the MLS determined either with⁵¹Cr or with¹¹¹In were lowered but the difference was not statistically significant, neither for group 1 (18.6 h vs 17.3 h;P>0.2) nor for group 2 (94.7 vs 122.3 h;P>0.2). The correlation between MLS and platelet counts in patients with AITP was significant for both labels (P<0.001). The 15 min recovery tended to be higher with¹¹¹In in all groups, but the difference was significant (P<0.05) only for group 1. The sequestration sites were similar with both labels. We conclude that, contrary to previous reports, platelet survival studies yield similar results with both the⁵¹Cr and¹¹¹In methods. Due to its distinct advantages¹¹¹In is the label of choice for investigation of platelet kinetics.Abbreviations ACD-A Anticoagulant citrate dextrose solution A: sodium citrate 2.2 g; citric acid 730 mg; dextrose 2.45 g and water for injections to 100 ml - AITP autoimmune thrombocytopenic purpura - ICSH International Committee for Standardization in Haematology - MLS mean life span Supported by the Deutsche Forschungsgemeinschaft (Mu 277/9-4)     

Electrophysiological study of transport systems in isolated perfused pancreatic ducts: properties of the basolateral membrane

In order to study the mechanism of pancreatic HCO ₃ ^– transport, a perfused preparation of isolated intra-and interlobular ducts (i.d. 20–40 m) of rat pancreas was developed. Responses of the epithelium to changes in the bath ionic concentration and to addition of transport inhibitors was monitored by electrophysiological techniques. In this report some properties of the basolateral membrane of pancreatic duct cells are described. The transepithelial potential difference (PD_te) in ducts bathed in HCO ₃ ^– -free and HCO ₃ ^– -containing solution was –0.8 and –2.6 mV, respectively. The equivalent short circuit current (I_sc) under similar conditions was 26 and 50 A·cm^–2. The specific transepithelial resistance (R_te) was 88 cm². In control solutions the PD across the basolateral membrane (PD_bl) was –63±1 mV (n=314). Ouabain (3 mmol/l) depolarized PD_bl by 4.8±1.1 mV (n=6) within less than 10 s. When the bath K⁺ concentration was increased from 5 to 20 mmol/l, PD_bl depolarized by 15.9±0.9 mV (n=50). The same K⁺ concentration step had no effect on PD_bl if the ducts were exposed to Ba²⁺, a K⁺ channel blocker. Application of Ba²⁺ (1 mmol/l) alone depolarized PD_bl by 26.4±1.4 mV (n=19), while another K⁺ channel blocker TEA⁺ (50 mmol/l) depolarized PD_bl only by 7.7±2.0 mV (n=9). Addition of amiloride (1 mmol/l) to the bath caused 3–4 mV depolarization of PD_bl. Furosemide (0.1 mmol/l) and SITS (0.1 mmol/l) had no effect on PD_bl. An increase in the bath HCO ₃ ^– concentration from 0 to 25 mmol/l produced fast and sustained depolarization of PD_bl by 8.5±1.0 mV (n=149). It was investigated whether the effect of HCO ₃ ^– was due to a Na⁺⁺-dependent transport mechanism on the basolateral membrane, where the ion complex transferred into the cell would be positively charged, or whether it was due to decreased K⁺ conductance caused by lowered intracellular pH. Experiments showed that the HCO ₃ ^– effect was present even when the bath Na⁺ concentration was reduced to a nominal value of 0 mmol/l. Similarly, the HCO ₃ ^– effect remained unchanged after Ba²⁺ (5 mmol/l) was added to the bath. The results indicate that on the basolateral membrane of duct cells there is a ouabain sensitive (Na⁺+K⁺)-ATPase, a Ba²⁺ sensitive K⁺ conductance and an amiloride sensitive Na⁺/H⁺ antiport. The HCO ₃ ^– effect on PD_bl is most likely due to rheogenic anion exit across the luminal membrane.     

The clinicopathologic and prognostic significance of CD44+/CD24(-/low) and CD44-/CD24+ tumor cells in invasive breast carcinomas

Cells with distinct phenotypes and stem cell-like properties have been reported to exist in breast cancer. The aim of the present study was to investigate the clinicopathologic and prognostic significance of the CD44(+)/CD24(-/low) and CD44(-)/CD24(+) tumor phenotypes' prevalence. Double immunohistochemistry was applied on a series of 155 paraffin-embedded breast tissue specimens to detect CD44 and CD24. Evaluation of the phenotypes was performed by image analysis. The prevalence of CD44(+)/CD24(-/low) and CD44(-)/CD24(+) tumor cells was 58.7% and 82.6%, respectively. The dominance of the CD44(+)/CD24(-/low) tumor cells was inversely associated with lymph node metastasis (P = .019) and tended to inversely associate with the stage of the disease (P = .068). Moreover, the prevalence of CD44(+)/CD24(-/low) was found to exert no significant impact on patients' prognosis although it displayed a tendency toward an increase in disease-free survival (P = .074). On the other hand, the prevalence of CD44(-)/CD24(+) tumor cells was found to have no clinicopathologic significance. However, it was found to exert an unfavorable impact on both relapse-free (P = .009) and overall survival (P = .046) of the patients with breast carcinomas of intermediate differentiation (grade 2). In breast tissue, CD44(+)/CD24(-/low) tumor cells seem to be associated with lack of lymph node metastasis and a tendency toward an increase of the relapse-free survival of the patients. On the contrary, tumor cells with the phenotype CD44(-)/CD24(+) seem to identify patients with worse disease-free and overall survival within the group of intermediate-grade differentiation patients whose prognosis is difficult to assess.     

Regulatory CD4+CD25+ T-cells are Controlled by Multiple Pathways at Multiple Levels

CD4⁺CD25⁺ regulatory T-cells (Treg cells) are an important subset of T-cells that functions to negatively control immune responses to self or non-self antigens. Depletion of CD4⁺CD25⁺ Treg cells leads to the occurrence of lymphoproliferative autoimmune diseases in animals and humans. Therefore, CD4⁺CD25⁺ Treg cells must be tightly regulated in the physiologic situation. In this article, we try to summarize the regulating pathways of the development, survival, and function of CD4⁺CD25⁺ Treg cells at multiple levels and multiple pathways, including the dendritic cells, costimulatory signals, cytokines, as well as intracellular signals.