Biomedical subjects

N Yang

Publications and source records attributed to N Yang.

At least 73 records · Page 4Linked to original sources

PubMed1998

De novo glomerular osteopontin expression in rat crescentic glomerulonephritis.

Osteopontin (OPN) is a secreted acidic glycoprotein that has potent monocyte chemoattractant and adhesive properties. Up-regulation of tubular OPN expression is thought to promote interstitial macrophage infiltration in experimental nephritis; however, the role of OPN in glomerular lesions, particularly crescent formation, is unknown. The present study used Northern blotting, in situ hybridization and immunohistochemistry to examine OPN expression in a rat model of accelerated anti-GBM glomerulonephritis. Osteopontin mRNA and protein is expressed by some parietal epithelial cells, thick ascending limbs of Henle and medullary tubules and collecting ducts in normal rat kidney. De novo OPN mRNA and protein expression was evident in glomerular visceral and parietal epithelial cells in anti-GBM glomerulonephritis. Glomerular OPN expression preceded and correlated with macrophage infiltration in the development of hypercellularity, focal and segmental lesions and, notably, crescent formation. There was marked up-regulation of OPN expression by tubular epithelial cells that also preceded and correlated with interstitial macrophage (r = 0.93, P < 0.001) and T-cell infiltration (r = 0.85, P < 0.001). Both glomerular and tubular OPN expression correlated significantly with proteinuria (P < 0.001) and a reduction in creatinine clearance (P < 0.01). In addition, double immunohistochemistry showed co-expression of osteopontin and one of its ligands, CD44, in intrinsic renal cells. CD44 and OPN expression by parietal epithelial cells was evident in crescent formation, while virtually all OPN-positive tubules expressed CD44. Infiltrating macrophages and T-cells were CD44-positive, but only a small proportion of T-cells and few macrophages showed OPN expression. Interestingly, strong OPN mRNA and protein expression was seen in macrophage multinucleated giant cells. In summary, this study suggests that OPN promotes macrophage and T-cell infiltration in the development of renal lesions in rat anti-GBM glomerulonephritis, including glomerular crescent and multinucleated giant cell formation.

Animals↗

PubMed1998

Local macrophage proliferation in human glomerulonephritis.

BACKGROUND: Local macrophage proliferation has been described in several animal models of glomerulonephritis (GN), but its significance in human disease is unknown. METHODS: Double immunostaining for CD68 and the proliferating cell nuclear antigen (PCNA) was used to identify macrophage proliferation in 84 biopsies from a variety of glomerulonephridities. RESULTS: A small resident population of glomerular and interstitial CD68+ macrophages was identified in normal human kidney, of which only 1 to 2% showed evidence of proliferation on the basis of PCNA expression. A mild macrophage infiltrate, with only occasional proliferating macrophages, was seen in the less aggressive forms of GN (minimal change disease, non-IgA mesangioproliferative GN and IgA nephropathy). This was in sharp contrast to the more aggressive forms of disease (lupus class IV, vasculitis-associated GN, crescentic GN and mesangiocapillary proliferative GN), in which the prominent macrophage infiltrates contained many proliferating macrophages, accounting for 28 to 47% of the total macrophage population. Macrophage proliferation was largely restricted to areas of severe tissue damage (glomerular segmental proliferative lesions, crescents and foci of tubulointerstitial damage), suggesting that local proliferation is a mechanism for amplifying macrophage-mediated injury. Glomerular and interstitial macrophage proliferation gave a significant correlation with loss of renal function (P < 0.0001) and histologic lesions (P < 0.0001), but not with proteinuria. Interstitial T-cell proliferation also gave a significant correlation with loss of renal function and histologic damage, even though proliferation within the T-cell population was much lower than in the macrophage population. CONCLUSIONS: This study demonstrates that macrophage proliferation is a feature of the more aggressive forms of human GN. Local proliferation may be an important mechanism for amplifying macrophage-mediated renal injury. In addition, the degree of local macrophage proliferation may be a useful diagnostic and prognostic indicator for human GN.

Adolescent↗

PubMed1998

Local macrophage and myofibroblast proliferation in progressive renal injury in the rat remnant kidney.

BACKGROUND: We have recently shown that blockade of angiotensin II activity inhibits local macrophage and myofibroblast proliferation in progressive non-immune renal injury in the rat remnant kidney. However, it is not known whether this local proliferation contributes to macrophage and myofibroblast accumulation and the development of renal injury. Therefore, we examined this issue in a detailed time-course study of the rat remnant kidney. METHODS: Groups of five rats were killed 4, 8,12 or 16 weeks after 5/6 subtotal nephrectomy (STNx) or a sham operation. Macrophage and myofibroblast proliferation was assessed by two-colour immunostaining for ED1+ macrophages or alpha-smooth muscle actin (alpha-SMA)-positive myofibroblasts with the proliferating cell nuclear antigen (PCNA) or bromodeoxyuridine. RESULTS: All parameters of renal function and histology remained normal in the sham-operated controls, and no macrophage or myofibroblast accumulation was evident. In contrast, prominent macrophage accumulation developed in both the glomerulus and tubulointerstitium in STNx animals, peaking at week 12. Many ED1+ macrophages showed PCNA expression, accounting for 19-34% of the total macrophage population. There was a highly significant correlation between proliferating macrophages and total macrophage accumulation in the glomerulus (r = 0.82, P < 0.0001) and tubulointerstitium (r = 0.70, P < 0.001). Macrophage proliferation was largely restricted to focal areas of renal damage, such as glomerular segmental lesions and severe tubulointerstitial damage. Also, the subpopulation of proliferating macrophages gave a highly significant correlation with loss of renal function, proteinuria, and glomerular and tubulointerstitial lesions. In addition, many alpha-SMA myofibroblasts were evident within expanded mesangial areas and the tubulointerstitium following STNx. Interestingly, active lesions contained many large alpha-SMA+ cells double-stained for PCNA, accounting for 24-29% of total myofibroblasts. There was a highly significant correlation between the number of proliferating myofibroblasts and total myofibroblast accumulation during the evolution of this disease, and both populations correlated with progressive renal injury. CONCLUSIONS: This study has shown that local proliferation is an important mechanism in both macrophage and myofibroblast accumulation during the development of renal injury in the rat remnant kidney. In addition, local macrophage proliferation is postulated as a mechanism for amplifying kidney damage in nonimmune renal injury.

Actins↗

PubMed1998

Forty generations of bidirectional selection for mating frequency in male Japanese quail.

A bidirectional replicated selection experiment for high (H1 and H2) or low (L1 and L2) cumulative number of complete matings (CNCM) in male Japanese quail was conducted for 40 generations. In the S32 generation, a subline was taken from each selected line and selection was relaxed. In the randombred control line (C), CNCM and unselected traits changed significantly over generations. Means of the selected lines were adjusted each generation for deviations from the control means. After 40 generations of selection, there was a 21-fold difference in CNCM (59.4 vs 2.8) between Lines H1 and L1. Whereas means increased and variation decreased in the high lines, means decreased and variation increased in the low lines. Regressions of mean CNCM on generation of Lines H1, H2, L1, and L2 were 1.15 +/- 0.08, 0.61 +/- 0.08, -0.26 +/- 0.04, and -0.34 +/- 0.03, respectively. Although responses to selection were observed throughout the 40 generations in Line H1, the low lines appeared to have reached a limit to selection after the S30 generation. Relaxed lines provided supporting evidence for this conclusion. Mean CNCM decreased in the relaxed high lines to that of the control, whereas the low relaxed lines remained at the same level as their corresponding selected lines. Line H2 went into extinction in the 37th generation as a result of reduced fitness. Realized heritabilities of CNCM were 0.09, 0.07, 0.06, and -0.15 in Lines H1, H2, L1, and L2, respectively. As correlated responses to the selection, male quail in the high lines were heavier, exhibited greater relative aggressiveness, and had larger cloacal glands than those of the control and low lines.

Animals↗

PubMed1998

Reversal of established rat crescentic glomerulonephritis by blockade of macrophage migration inhibitory factor (MIF): potential role of MIF in regulating glucocorticoid production.

Macrophage migration inhibitory factor (MIF) is a potent pro-inflammatory cytokine that also counter-regulates glucocorticoid action. We investigated whether immunoneutralization of MIF could reverse established experimental crescentic glomerulonephritis and if this treatment could modulate endogenous glucocorticoid levels. Accelerated anti-GBM glomerulonephritis was induced in six littermate pairs of rats. Once crescentic disease was established on day 7, one animal in each pair was given a daily injection of neutralizing anti-MIF antibody (Ab) or irrelevant isotype control Ab for 14 days and then killed on day 21. In addition, a group of 6 animals was killed on day 7 of disease without any treatment. Animals receiving the control Ab exhibited a rapidly progressive glomerulonephritis with severe renal injury (proteinuria), loss of renal function (creatinine clearance), anemia, and marked histologic damage (including glomerular crescent formation), compared with animals killed on day 7 without treatment. In contrast, anti-MIF Ab treatment partially reversed the disease by restoring normal renal function and reducing histological damage compared with untreated animals killed on day 7 (p < 0.05). Interestingly, anti-MIF Ab treatment also prevented severe anemia (p < 0.05). Reversal of disease was associated with a significant reduction in leukocyte infiltration and activation and renal interleukin-1 (IL-1) production. Importantly, anti-MIF Ab treatment caused a significant increase in endogenous serum corticosterone levels, which correlated with the reversal of disease parameters. In conclusion, this study has demonstrated that blocking MIF activity can partially reverse established crescentic glomerulonephritis and suggests that MIF operates by both enhancing the cellular immune response and suppressing the endogenous anti-inflammatory glucocorticoid response.

Animals↗

PubMed1998

[Alanyl-glutamine dipeptides protected the liver function by increasing the hepatic glutathione].

OBJECTIVE: Glutathione (GSH) is a major antioxidant which protects hepatic tissues from free radical injury. Alanyl-Glutamine (ALA-GLN) proved to be a precursor of GSH synthesis, was used to investigate the relationship to GSH biosynthesis which may be effective for hepatic protection. METHODS: 20 male Wistar rats were randomly divided into two groups receiving standard parenteral nutrition (STD) supplemented with or without ALA-GLN for 7 days. At 5th day 5-fluorouracil (5-FU) was injected peritoneally, the blood samples for GSH, GSSG, ALT (sGPT), AKP and TBilli tests were measured after 4-8 h. RESULTS: The concentration measurements were significantly different in ALA-GLN group compared with STD animals in serum GLN (687.3 +/- 49.8) vs (504.9 +/- 38.6) mumol/L, P < 0.05), serum GSH (14.37 +/- 5.16) vs (7.08 +/- 3.16) mumol/L, P < 0.01) and in liver GSH content (6.86 +/- 2.46) vs (4.38 +/- 1.63) mumol/g liver tissue, P < 0.05). Rats in ALA-GLN group have lesser elevations in hepatic enzymes after 5-FU administration. CONCLUSIONS: The supplemented nutrition ALA-GLN protected the liver function through increasing the glutathione biosynthesis and preserving the glutathione stores of hepatic tissue.

Animals↗

PubMed1998

[Clinical study on effect of Chinese herbal medicine combined with hemodialysis in treating uremia].

OBJECTIVE: To elevate the efficacy of dialysis in uremia patients treated with maintaining hemodialysis. METHODS: Thirty patients of terminal stage uremia were treated with Tongmai oral liquid and rhubarb capsule combined with hemodialysis were observed and compared with a control group of 30 patients treated with hemodialysis alone. The therapeutic course of both groups was 1 month and the observation lasted for 3 months consecutively. RESULTS: The mean time urea nitrogen concentration, protein catabdic rate and the GS index (KT/V value) of the treated group were all better than those of the control group (P < 0.05). CONCLUSION: Uremia patients treated with Tongmai oral liquid and rhubarb capsule combined with hemodialysis were better in full utilization of hemodialysis and life quality of patients in comparison with patients treated with hemodialysis alone.

Adult↗

PubMed1998

[Esophageal insufflation test in secondary voice reconstruction].

OBJECTIVE: To restore the alaryengeal patients' enunciation function, reduce operative trauma, increase the accuracy of detecting the success of secondary voice restoration and evaluate objectively the practical clinical significance of esophageal insufflation test. METHODS: Esophageal insufflation test and secondary voice restoration were done for 45 alaryngeal patients, especially those with negative results of esophageal insufflation tests. RESULTS: It showed that the success rate of voice rehabilitation in patients with negative insufflation test but no intact pharyngeal constrictor was 94.7%(36/38), in those with positive insufflation test was 100%(7/7) uniformly. The postoperative pronounciation performance was better than that at the test time. CONCLUSION: Esophageal insufflation test is a useful procedure in predicting the postoperative pronunciation, but could be influenced by many factors, so it can not be used as the only test for estimating the postoperative pronunciation.

Adult↗

PubMed1998

A comparative study of transvaginal ultrasonography and pelvic arteriogram in assessment of patients with gestational trophoblastic tumour.

OBJECTIVE: To compare the reliability of transvaginal ultrasonography with pelvic arteriography in the assessment of patients with gestational trophoblastic disease. METHODS: Transvaginal ultrasonography was performed in 24 patients with gestational trophoblastic tumour. Within one week after ultrasound investigation, pelvic arteriography was carried out in each patient. Of 24 cases, 16 patients hadn't been treated by chemical reagent, 5 had accepted 2 to 5 courses of chemotherapy, and 3 had achieved complete remission before both investigations performed. RESULTS: In 3 patients with complete remission, 2 had no evidence of abnormal findings either on transvaginal ultrasonography or on pelvic arteriography, 1 showed intramyometrial lesions by both methods. In the remaining 21 patients, all demonstrated a abnormal uterine image, and 5 of them accompanied with the finding of parametrium metastatic signs by transvaginal ultrasonography; these abnormal results were confirmed by pelvic arteriographic imaging. However, in two cases without clinical and ultrasonic signs of parametrium metastasis, pelvic arteriography indicated the early metastasis of parametrium vessels. CONCLUSIONS: Even though it is difficult to predict the early parametrium metastasis in patients with gestational trophoblastic disease by B-ultrasonic investigation, our data would support the introduction of transvaginal ultrasonography in the diagnosis and evaluation of gestational trophoblastic tumour.

Angiography↗

PubMed1998

[Research and development of the surface EMG detection and analysis system].

To meet the requirement of developing myoelectric controller for a man-machine systems, a surface electromyography (EMG) detection and analysis system is discussed in this paper. This system consists of two parts--data acquisition hardware and signal processing software. In the surface EMG acquisition system both wire and wireless TELEMG are available. In the surface EMG processing system various methods are integrated such as the methods in time domain, the methods for frequency analysis, and also including some newly developed methods, e.g., wavelet transform and fractal analysis. This system provides a useful platform for research and development of the myoelectric controller that is widely used in rehabilitation engineering products.

Electromyography↗

PubMed1998

Mouse lactoferrin gene. Promoter-specific regulation by EGF and cDNA cloning of the EGF-response-element binding protein.

Expression of the lactoferrin gene in a variety of tissues is regulated differentially. We have previously demonstrated that the lactoferrin gene is regulated by estrogen and mitogen in mouse uterus. The mouse lactoferrin gene responded to forskolin, cAMP, TPA and EGF stimulation via two adjacent enhancer elements, the CRE and EGFRE and collectively referred to as the Mitogen Response Unit (MRU). We found that CRE is responsible for forskolin, cAMP and TPA whereas EGFRE is for EGF stimulation. We examined the minimal promoter and enhancer elements of the mouse lactoferrin gene that are required for EGF induced transcriptional activation. We found that the CRE and noncanonical TATA box (ATAAA) are the minimal promoter elements for basal activity of the CAT reporter construct, whereas, the EGFRE is needed for an additional activity induced by EGF in transiently transfected human endometrial carcinoma RL95-2 cells (RL95-2). The EGFRE, however, did not function in heterologous promoters (SV 40 and TK). Therefore, EGF-stimulated lactoferrin gene activity is promoter specific in RL95-2 cells. Mutation made at either elements or insertion of extra nucleotides between the two elements, severely affected EGF-stimulated activity. Nuclear protein prepared from RL95-2 cells protected the EGFRE, CRE and noncanonical TATA from DNAase I digestion in a footprinting analysis. Nuclear protein which interacted with the CRE were previously identified as API and CREB. In this study, we isolated a cDNA clone from an RL95-2 expression library that encodes the EGFRE binding protein. Partial sequence of the cDNA clone revealed 100% nucleotide identity with a GC-box binding protein, BTEB2. Protein-protein interaction among the transcription factors could fine-tune the mouse lactoferrin expression in various tissues.

Animals↗

PubMed1998

An improved, inexpensive method for the large-scale purification of human nerve growth factor.

Human nerve growth factor (hNGF) was purified to near homogeneity on a large scale from human term placenta with an improved and inexpensive method. The purification procedure included tissue homogenization, ultrafiltration and single CM-cellulose column chromatography. The purified hNGF was a 14.4-kDa protein with an isoelectric point of approximately 9.3. The specific activity of the purified hNGF was approximately 38000 units/mg, and the activity was completely inhibited by the monoclonal antibody against recombinant hNGF (rhNGF). Western-blot analysis showed that the purified hNGF could interact with the monoclonal antibody against rhNGF.

China↗

PubMed1997

Characterization of the alpha1B-adrenergic receptor gene promoter region and hypoxia regulatory elements in vascular smooth muscle.

We previously demonstrated that alpha1B-adrenergic receptor (AR) gene transcription, mRNA, and functionally coupled receptors increase during 3% O2 exposure in aorta, but not in vena cava smooth muscle cells (SMC). We report here that alpha1BAR mRNA also increases during hypoxia in liver and lung, but not heart and kidney. A single 2.7-kb alpha1BAR mRNA was detected in aorta and vena cava during normoxia and hypoxia. The alpha1BAR 5' flanking region was sequenced to -2,460 (relative to ATG +1). Transient transfection experiments identify the minimal promoter region between -270 and -143 and sequence between -270 and -248 that are required for transcription of the alpha1BAR gene in aorta and vena cava SMC during normoxia and hypoxia. An ATTAAA motif within this sequence specifically binds aorta, vena cava, and DDT1MF-2 nuclear proteins, and transcription primarily initiates downstream of this motif at approximately -160 in aorta SMC. Sequence between -837 and -273 conferred strong hypoxic induction of transcription in aorta, but not in vena cava SMC, whereas the cis-element for the transcription factor, hypoxia-inducible factor 1, conferred hypoxia-induced transcription in both aorta and vena cava SMC. These data identify sequence required for transcription of the alpha1BAR gene in vascular SMC and suggest the atypical TATA-box, ATTAAA, may mediate this transcription. Hypoxia-sensitive regions of the alpha1BAR gene also were identified that may confer the differential hypoxic increase in alpha1BAR gene transcription in aorta, but not in vena cava SMC.

Animals↗

PubMed1997

Human estrogen receptor-like 1 (ESRL1) gene: genomic organization, chromosomal localization, and promoter characterization.

Estrogen receptor-like 1a (ESRL1a; same as estrogen receptor-related orphan receptors, ERR1) belongs to a subfamily of the nuclear receptor superfamily. We have previously shown that human ESRL1a modulates estrogen responsiveness of the lactoferrin gene promoter in transiently transfected endometrial carcinoma RL95-2 cells. In this study, we cloned and characterized the human ESRL1 gene. Through the fluorescence in situ hybridization method, the ESRL1 gene was localized to the centromere region of chromosome 11q12. Partial sequencing, restriction mapping, and PCR analysis revealed that the ESRL1 gene consists of seven exons and is approximately 20 kb in length. We found that the smallest exon (exon 3) contains 117 bp and the largest exon (exon 7) has 1032 bp. The smallest intron (intron 5) is only 88 bp long and the largest intron (intron 2) is 8 kb long. All introns have the conserved GT and AG dinucleotides present at the donor and acceptor sites, respectively. Like the estrogen receptor, the highly conserved DNA-binding domain of hESRL1a is encoded by exon 2 and exon 3, and the intron/exon junctions (2 and 3) are well conserved between the two genes. Primer extension analysis revealed multiple transcription initiation start sites in human uterine (HeLa, HEC, and RL95-2) cell lines. However, one major initiation start site was found by RNase protection assay. The hESRL1a mRNA is differentially expressed in various human tissues. The nucleotide sequence adjacent to the transcription start sites of the ESRL1 lacks the typical TATA and CAAT boxes but is GC rich and contains 10 consensus Sp1-binding elements and two E boxes. The region that contains these transcription factor-binding elements showed a high level of promoter activity when transiently transfected into RL95-2 cells.

Blotting, Northern↗

PubMed1997

The pathogenic role of macrophage migration inhibitory factor in immunologically induced kidney disease in the rat.

Macrophage migration inhibitory factor (MIF) plays a pivotal role in the inflammatory response in endotoxemia and in the delayed-type hypersensitivity response, but its potential as a regulator of immunologically induced disease is unknown. We have addressed this issue by administering a neutralizing anti-MIF antibody in a rat model of immunologically induced crescentic anti-glomerular basement membrane (GBM) glomerulonephritis. Six individual experiments using paired inbred littermates were performed. Rats were primed with rabbit immunoglobulin on day -5 and then injection with rabbit anti-rat GBM serum on day 0. Pairs of animals were treated with anti-MIF or a control monoclonal antibody from the time of anti-GBM serum administration until being killed 14 d later. Control antibody-treated animals developed severe proteinuria and renal function impairment with severe histological damage due to marked leukocytic infiltration and activation within the kidney. In contrast, anti-MIF treatment substantially reduced proteinuria, prevented the loss of renal function, significantly reduced histological damage including glomerular crescent formation, and substantially inhibited renal leukocytic infiltration and activation (all P <0.001 compared with control treatment). Inhibition of renal disease by anti-MIF treatment was attributed to preventing the marked upregulation of interleukin-1beta, leukocyte adhesion molecules including intercellular adhesion molecule-1 and vascular cell adhesion molecule-1, and inducible nitric oxide synthase expression seen in the control antibody-treated animals. This inhibition of progressive renal injury was mirrored by the complete suppression of the skin delayed-type hypersensitivity response to the challenge antigen (rabbit IgG). Interestingly, anti-MIF treatment did not effect the secondary antibody response or immune deposition within the kidney, indicating that MIF participates in cellular-based immunity in this primed macrophage-dependent anti-GBM glomerulonephritis. In conclusion, this study has demonstrated a key regulatory role for MIF in the pathogenesis of immunologically induced kidney disease. These results argue that blocking MIF activity may be of benefit in the treatment of human rapidly progressive glomerulonephritis, and suggest that MIF may be important in immune-mediated disease generally.

Animals↗

PubMed1997

Inhibition of activated Ras-induced neuronal differentiation of PC12 cells by the LIM domain of LIM-kinase 1.

LIM-kinase 1 and 2 (LIMK1 and LIMK2) are members of a novel class of protein kinases with structures composed of two LIM motifs at the N-terminus and an unusual protein kinase domain at the C-terminus. The cellular functions of the LIMK family proteins have remained unknown. In the present study, we examined effects of LIMKs on neuronal differentiation of PC12 pheochromocytoma cells. Transient expression analyses revealed that LIMK1, in itself, had no apparent effect on PC12 cells, but the oncogenic Ras-induced differentiation of PC12 cells was notably inhibited by co-expression with LIMK1 or LIMK2. A mutant of LIMK1 lacking a protein kinase domain (delta K) similarly inhibited Ras-induced differentiation of PC12 cells, but a mutant lacking a LIM domain (delta LIM) failed to do so, indicating that a LIM domain but not a protein kinase domain is required for the inhibitory activity. This notion was further supported by the finding that mutation, changing conserved cysteines involved in zinc coordination to glycines in both of two LIM motifs, abolished the inhibitory activity of delta K. Additionally, we also found that the constitutively activated MAP kinase kinase (MAPKK)-induced differentiation of PC12 cells was inhibited by co-expression with delta K. Furthermore, AK did not inhibit the kinase activity of MAP kinase (MAPK) stimulated by MAPKK, when co-expressed in COS7 cells. These findings suggest that LIMK1 inhibits neuronal differentiation of PC12 cells, through its LIM domain and by interfering with events downstream of MAPK activation.

Animals↗

PubMed1997

Probing the outer vestibule of a sodium channel voltage sensor.

The second and third basic residues of the S4 segment of domain 4 (D4:R2 and D4:R3) of the human skeletal muscle Na+ channel are known to be translocated from a cytoplasmic to an extracellular position during depolarization. Accessibilities of individual S4 residues were assayed by alteration of inactivation kinetics during modification of cysteine mutants by hydrophilic methanethiosulfonate reagents. The voltage dependences of the reaction rates are identical for extracellular application of cationic methanethiosulfonate-ethyltrimethylammonium (MTSET) and anionic methanethiosulfonate-ethylsulfonate (MTSES), suggesting that D4:R3C is situated outside the membrane electric field at depolarized voltages. The absolute rate of R3C modification is 281-fold greater for MTSET than for MTSES, however, suggesting that at depolarized voltages this S4 thiol resides in a negatively charged hydrophilic crevice. The two hydrophobic residues between D4:R2C and D4:R3C in the primary sequence (L1452 and A1453) are not externally exposed at any voltage. An alpha-helical representation of D4/S4 shows that the basic residues D4:R2 and D4:R3 are on the face opposite that of L1452 and A1453. We propose that in the depolarized conformation, the hydrophobic face of this portion of D4/S4 remains in contact with a hydrophobic region of the extracellular vestibule of the S4 channel.

Alkanesulfonates↗

PubMed1997

Intrinsic renal cells are the major source of interleukin-1 beta synthesis in normal and diseased rat kidney.

BACKGROUND: A number of studies have demonstrated a pathological role for interleukin-1 (IL-1) in experimental models of glomerulonephritis, but the cellular pattern of renal IL-1 production remains poorly characterized. The aim of this study, therefore, was to identify the cell types expressing IL-1 in normal and diseased rat kidney. METHODS: Renal IL-1 beta expression was examined in normal rats and during a 21-day time course of rat accelerated anti-GBM glomerulonephritis by northern blotting, in situ hybridization and double immunohistochemistry. RESULTS: Interleukin-1 beta mRNA expression was readily detectable in normal rat kidney by northern blot analysis and in situ hybridization. Immunohistochemistry staining demonstrated constitutive IL-1 beta expression by glomerular endothelial cells and cortical tubular epithelial cells. There was a marked increase in whole kidney IL-1 beta mRNA in rat anti-GBM glomerulonephritis. Glomerular IL-1 beta immunostaining was upregulated, being expressed by podocytes, mesangial cells and infiltrating macrophages, and was particularly prominent within glomerular crescents. Double staining with the ED1 antibody showed IL-1 beta expression in up to 13% of glomerular macrophages, whereas 48% of macrophages within crescents stained for IL-1 beta. However, the most marked increase in IL-1 beta expression was seen in cortical tubular epithelial cells, particularly in areas of tubular damage. In situ hybridization confirmed that tubular IL-1 beta staining was due to local cytokine synthesis rather than protein absorption. CONCLUSIONS: This study has identified constitutive IL-1 beta expression by glomerular endothelium and tubular epithelial cells in normal rat kidney. In addition, the marked upregulation of IL-1 beta expression by intrinsic glomerular cells and tubules in rat anti-GBM disease suggests an important role for these cells in IL-1 dependent crescent formation and tubulointerstitial injury.

Animals↗