Injury and protection of spiral ganglion neurons.

ABSTRACT: Cochlear spiral ganglion neurons (SGNs) are bipolar ganglion cells and are the first neurons in the auditory transduction pathway. They transmit complex acoustic information from hair cells to second-order sensory neurons in the cochlear nucleus for sound processing. Injury to SGNs causes largely irreversible hearing impairment because these neurons are highly differentiated cells and cannot regenerate, making treatment of sensorineural hearing loss (SNHL) arising from SGN injury difficult. When exposed to ototoxic drugs or damaging levels of noise or when there is loss of neurotrophic factors (NTFs), aging, and presence of other factors, SGNs can be irreversibly damaged, resulting in SNHL. It has been found that NTFs and stem cells can induce regeneration among dead spiral ganglion cells. In this paper, we summarized the present knowledge regarding injury, protection, and regeneration of SGNs.

Bioinformatics analysis and verification of key candidate genes influencing the pathogenesis of chronic rhinosinusitis with nasal polyps.

OBJECTIVES: Chronic rhinosinusitis (CRS) with nasal polyps (CRSwNP) is a prominent public health issue. Furthermore, the prognosis of eosinophilic CRSwNP is poor, with a high recurrence rate. The underlying molecular mechanisms of eosinophilic CRSwNP remain unclear. Therefore, in this study, we sought to determine the crucial genes underlying eosinophil infiltration in eosinophilic CRSwNP pathogenesis. METHODS: We used the Gene Expression Omnibus database (GEO) (GSE36830 and GSE23552 datasets) to mine gene expression profiles of CRSwNP patients and normal subjects. Differentially expressed genes (DEGs) between normal and CRSwNP tissues were identified and subjected to the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analyses. Co-expression networks were established using a weighted gene co-expression network analysis (WGCNA) and single-sample gene set enrichment analysis (GSEA). Protein-protein interaction networks were developed to detect functional protein modules. Based on the common DEGs, candidate miRNAs and related lncRNAs were predicted using the mirTarBase and StarBase databases. Finally, we generated immune cell subtypes of CRSwNP. RESULTS: A total of 146 DEGs were identified. Of these, 131 genes were upregulated, whereas 15 were downregulated. GO analysis indicated that DEGs primarily participated in leukocyte chemotaxis and migration as well as cell chemotaxis. KEGG pathway analysis suggested that DEGs participated in the interactions between cytokines and viral proteins, osteoclast differentiation, and cytokine-cytokine receptor interactions. Real-time quantitative polymerase chain reaction analysis showed that Complement C5a Receptor 1 (C5AR1), C-C Motif Chemokine Receptor 3 (CCR3), Complement C3a Receptor 1 (C3AR1), and C-C Motif Chemokine Ligand 13 (CCL13) expression levels were significantly upregulated in nasal polyps, whereas C-C Motif Chemokine Ligand 4 (CCL4) expression levels were significantly downregulated. CONCLUSIONS: The candidate genes identified in this study may influence the activation and accumulation of eosinophils, cell chemotaxis, and inflammatory responses, thereby potentially representing molecular targets for future studies of CRSwNP.

Heterozygous MYH9 Mutations in 2 Children With Cochlear Nerve Canal Stenosis.

MYH9 is a gene that encodes for a subunit of the myosin heavy chain IIA protein. Mutations in MYH9 are associated with hematologic abnormalities, renal dysfunction, and hearing loss. Bony cochlear nerve canal stenosis (CNCS), which is diagnosed on computed tomography (CT) imaging, has been associated with congenital deafness, cochlear nerve aplasia/hypoplasia, and inner ear malformations. We report two cases of CNCS presenting with profound congenital hearing loss whom we diagnosed with mutations in MYH9 and discuss the genotype-phenotype association and implications for management.

Clinical characteristics in unilateral cochlear nerve canal stenosis.

OBJECTIVE: To characterize the clinical features of patients with congenital hearing loss and unilateral cochlear nerve canal stenosis (CNCS). METHODS: A retrospective review of 12 patients with unilateral CNCS diagnosed between January 2018 and December 2019 at a tertiary referral hospital was performed. RESULTS: Of the 12 patients identified, there were 6 males and 6 females. All patients presented with hearing loss, with no other chief complaints. Two patients had accessory auricles. Eleven patients had a severe to profound sensorineural hearing loss on the affected side, while 1 patient had an isolated high-frequency hearing loss. Nine patients demonstrated atresia of the cochlear nerve canal (CNC), while three patients had a stenotic, but patent, CNC. CONCLUSION: Prompt radiologic diagnosis of patients with unilateral CNCS is important for patient counseling and appropriate rehabilitation.

Auditory and speech performance after unilateral cochlear implantation for cochlear nerve canal stenosis.

To explore the correlation between the width of the bony cochlear nerve canal (CNC) and long-term auditory rehabilitation after unilateral cochlear implantation (CI) in pediatric patients with congenital deafness and bilateral cochlear nerve canal stenosis (CNCS). A retrospective review was performed on 10 patients with bilateral CNCS and bilateral congenital profound hearing loss who each underwent unilateral cochlear implantation. The width of the CNC was determined on computed tomography (CT) imaging and following CI, auditory and speech performance following CI were graded using categories of auditory performance (CAP), speech intelligibility rating (SIR), and the meaningful auditory integration scale (MAIS) at 24 months following implantation. No correlation was noted between CAP score and CNCS at 24 months post CI (P > .05). A positive correlation was noted between SIR score and CNC width (ρ = .81, P < .05). Similarly, a positive correlation was noted between MAIS and CNC width (ρ = .71, P < .05). The width of the CNC in patients with CNCS is positively correlated with some long-term auditory and speech outcomes after CI.

Recording of electrocochleography from the facial nerve canal in mice.

BACKGROUND: The ever-expanding arsenal of genetically modified mice has created experimental models for studying various mechanisms of deafness. Electrocochleography (ECochG) is a recording technique of cochlear potentials evoked by sound stimulation, which was widely used to evaluate the cochlear hearing function. However, there is currently a lack of information on long-term recording technology of ECochG in mice. NEW METHOD: We describe in detail the surgical procedure of implanting electrode into the facial nerve canal in C57BL/6J mice for ECochG recording. The results of ECochG recorded by electrode in the facial nerve canal were compared with ECochG guided by electrode on the round window niche. RESULTS: The surgical method of inserting the electrode into the facial nerve canal is relatively simple and can be completed within 15 min. The electrode inserted into the elongated facial nerve canal is stable and close to the auditory nerve trunk, so it is conducive to long-term auditory function monitoring. Hence, the ECochG guided by the electrode from the facial nerve canal can maintain a stable response for more than two weeks. In contrast, the ECochG guided by the electrode in the round window niche can only be maintained for a maximum of 20 min. COMPARISON WITH EXISTING METHODS: In mice, existing recording techniques of ECochG from round window niche is limited by conductive hearing loss due to middle ear effusion or surgical damage. CONCLUSIONS: ECochG recording from the facial nerve canal is suitable for long-term recording in mice. This electrode approach provides a repeatable and reliable measurement of ECochG.

Cochlear Nerve Canal Stenosis: Association With MYH14 and MYH9 Genes.

The bony cochlear nerve canal transmits the cochlear nerve as it passes from the fundus of the internal auditory canal to the cochlea. Stenosis of the cochlear nerve canal, defined as a diameter less than 1.0 mm in transverse diameter, is associated with inner ear anomalies and severe to profound congenital hearing loss. We describe an 11-month-old infant with nonsyndromic congenital sensorineural hearing loss with cochlear nerve canal stenosis. Next-generation sequencing revealed heterozygous mutations in MYH9 and MYH14, encoding for the inner ear proteins myosin heavy chain IIA and IIC. The patient's hearing was rehabilitated with bilateral cochlear implantation.

[Prediction of facial nerve fronting with temporal bone high resolution computed tomograph].

Objective:To report a new CT measurement to predict the exposure of round window niche during facial nerve recess surgery. Methods:Forty CT scans were measured by conventional facial-round window line and limiting facial-round window line method, and compared with the exposure of round window niche after the opening of the facial recess. Results:There was no significant difference in the sensitivity and specificity of the two CT measurement methods to predict the exposure degree of the round window niche, and the Kappa consistency test of the two methods also showed a strong correlation. Conclusion:The new measurement method can predict the exposure of the round window niche pre-operation, and in practice, it can predict the maximum exposure of the round window niche.

[Clinical analysis of 8 cases of delayed facial paralysis after middle ear surgery].

Objective:To investigate the possible causes, prevention, treatment and recovery of delayed facial paralysis after middle ear surgery. Method:A retrospectively analysis of the data of 8 patients with delayed facial paralysis after middle ear surgery under general anesthesia, including one case of tympanoplasty （type Ⅰ） , one case of tympanotomy+tympanoplasty c（type Ⅰ） , one case ofepitympanotomy+reconstruction of attic lateral wall+tympanoplasty （type Ⅱ ）, four cases of canal wall down+tympanoplasty （ type Ⅱ） and one case of canal wall up mastoidectomy. After discovering the facial paralysis, the stuffing in the surgical cavity was released and removed for all patients immediately. Necessary stuffing was replaced by dexamethasone gauze （not press hardl）. Meanwhile, patients took methylprednisolone orally and were intramuscularly injected with mecobalamine. Result:Among eight patients, there were six patients with horizontal exposure of facial nerve and two patients with pyramis exposure of facial nerve. There was one case, five cases and two cases of delayed facial paralysis at five days, one week and two weeks after operation respectively. There were six patients suffering from the facial paralysis of HB Ⅱ grade and two patients suffering from the facial paralysis of HB Ⅲ grade. Four patients recovered four weeks and the remaining four patients returned to normal six weeks after surgery. Conclusion:Delayed facial paralysis is one of complications of the middle ear surgery, and most of patients can recover completely after conservative treatment.

[Clinical features and surgical treatment of external auditory canal cholesteatoma in 149 cases].

Objective:The aim of this study is to explore the clinical characteristics, surgical management and treatment results of type Ⅰto type Ⅳ external auditory canal cholesteatoma（EACC）. Method:One hundred and forty-nine patients（150 ears） with EACC underwent different surgical approach according to the classification of EACC and the lesion range: ① 44 ears: external auditory canal lesion resection with or without reconstruction of external auditory canal ② 23 ears: external auditory canal lesion resection with reconstruction of external auditory canal and the tympanoplasty（TypesⅠto Ⅲ）; ③ 32 ears: external auditory canal lesion resection with reconstruction of external auditory canal and modified mastoidectomy and reconstruction of the posterior wall of external auditory canal; ④ 28 ears: external auditory canal lesion resection with reconstruction of external auditory canal and tympanoplasty（Types Ⅰ to Ⅲ） and modified mastoidectomy and reconstruction of the posterior wall of external auditory canal; ⑤12 ears: canal wall down mastoidectomy （CWD） with plasty of the cavity of auricular concha; ⑥ 11 ears: epitympanum dectomy and reconstruction with tympanoplasty. Result:In the 150 ears, there were 38 ears classified as Type Ⅰ, 52 as Type Ⅱ, 58 as Type Ⅲ and 2 as Type Ⅳ based on the Shin classification. All patients were followed up for more than half a year. The postoperative outcomes were satisfactory with low rate of cholesteatoma recurrence and the hearing was improved to varying degrees. Conclusion:Base on the variety of lesions, the surgical treatment method of choice depends on the extent of the lesion. Effective postoperative follow-up can reduce recurrence and avoid the second operation.

LXRα Regulates ChREBPα Transactivity in a Target Gene-Specific Manner through an Agonist-Modulated LBD-LID Interaction.

The cholesterol-sensing nuclear receptor liver X receptor (LXR) and the glucose-sensing transcription factor carbohydrate responsive element-binding protein (ChREBP) are central players in regulating glucose and lipid metabolism in the liver. More knowledge of their mechanistic interplay is needed to understand their role in pathological conditions like fatty liver disease and insulin resistance. In the current study, LXR and ChREBP co-occupancy was examined by analyzing ChIP-seq datasets from mice livers. LXR and ChREBP interaction was determined by Co-immunoprecipitation (CoIP) and their transactivity was assessed by real-time quantitative polymerase chain reaction (qPCR) of target genes and gene reporter assays. Chromatin binding capacity was determined by ChIP-qPCR assays. Our data show that LXRα and ChREBPα interact physically and show a high co-occupancy at regulatory regions in the mouse genome. LXRα co-activates ChREBPα and regulates ChREBP-specific target genes in vitro and in vivo. This co-activation is dependent on functional recognition elements for ChREBP but not for LXR, indicating that ChREBPα recruits LXRα to chromatin in trans. The two factors interact via their key activation domains; the low glucose inhibitory domain (LID) of ChREBPα and the ligand-binding domain (LBD) of LXRα. While unliganded LXRα co-activates ChREBPα, ligand-bound LXRα surprisingly represses ChREBPα activity on ChREBP-specific target genes. Mechanistically, this is due to a destabilized LXRα:ChREBPα interaction, leading to reduced ChREBP-binding to chromatin and restricted activation of glycolytic and lipogenic target genes. This ligand-driven molecular switch highlights an unappreciated role of LXRα in responding to nutritional cues that was overlooked due to LXR lipogenesis-promoting function.

OGT (O-GlcNAc Transferase) Selectively Modifies Multiple Residues Unique to Lamin A.

The LMNA gene encodes lamins A and C with key roles in nuclear structure, signaling, gene regulation, and genome integrity. Mutations in LMNA cause over 12 diseases ('laminopathies'). Lamins A and C are identical for their first 566 residues. However, they form separate filaments in vivo, with apparently distinct roles. We report that lamin A is ß-O-linked N-acetylglucosamine-(O-GlcNAc)-modified in human hepatoma (Huh7) cells and in mouse liver. In vitro assays with purified O-GlcNAc transferase (OGT) enzyme showed robust O-GlcNAcylation of recombinant mature lamin A tails (residues 385⁻646), with no detectable modification of lamin B1, lamin C, or 'progerin' (Δ50) tails. Using mass spectrometry, we identified 11 O-GlcNAc sites in a 'sweet spot' unique to lamin A, with up to seven sugars per peptide. Most sites were unpredicted by current algorithms. Double-mutant (S612A/T643A) lamin A tails were still robustly O-GlcNAc-modified at seven sites. By contrast, O-GlcNAcylation was undetectable on tails bearing deletion Δ50, which causes Hutchinson⁻Gilford progeria syndrome, and greatly reduced by deletion Δ35. We conclude that residues deleted in progeria are required for substrate recognition and/or modification by OGT in vitro. Interestingly, deletion Δ35, which does not remove the majority of identified O-GlcNAc sites, does remove potential OGT-association motifs (lamin A residues 622⁻625 and 639⁻645) homologous to that in mouse Tet1. These biochemical results are significant because they identify a novel molecular pathway that may profoundly influence lamin A function. The hypothesis that lamin A is selectively regulated by OGT warrants future testing in vivo, along with two predictions: genetic variants may contribute to disease by perturbing OGT-dependent regulation, and nutrient or other stresses might cause OGT to misregulate wildtype lamin A.

O-GlcNAc site-mapping of liver X receptor-α and O-GlcNAc transferase.

The Liver X Receptor α (LXRα) belongs to the nuclear receptor superfamily and plays an essential role in regulating cholesterol, lipid and glucose metabolism and inflammatory responses. We have previously shown that LXRα is post-translationally modified by O-linked ß-N-acetyl-glucosamine (O-GlcNAc) with increased transcriptional activity. Moreover, we showed that LXRα associates with O-GlcNAc transferase (OGT) in vitro and in vivo in mouse liver. In this study, we report that human LXRα is O-GlcNAc modified in its N-terminal domain (NTD) by identifying a specific O-GlcNAc site S49 and a novel O-GlcNAc modified peptide 20LWKPGAQDASSQAQGGSSCILRE42. However, O-GlcNAc site-mutations did not modulate LXRα transactivation of selected target gene promoters in vitro. Peptide array and co-immunoprecipitation assays demonstrate that LXRα interacts with OGT in its NTD and ligand-binding domain (LBD) in a ligand-independent fashion. Moreover, we map two new O-GlcNAc sites in the longest OGT isoform (ncOGT): S437 in the tetratricopeptide repeat (TPR) 13 domain and T1043 in the far C-terminus, and a new O-GlcNAc modified peptide (amino acids 826-832) in the intervening region (Int-D) within the catalytic domain. We also map four new O-GlcNAc sites in the short isoform sOGT: S391, T393, S399 and S437 in the TPRs 11-13 domain. Future studies will reveal the biological role of identified O-GlcNAc sites in LXRα and OGT.

Characteristics of electrically evoked auditory brainstem responses in patients with cochlear nerve canal stenosis receiving cochlear implants.

OBJECTIVE: To explore the characteristics of the electrically evoked auditory brainstem responses (EABR) in children with cochlear nerve canal stenosis (CNCs) following cochlear implantation (CI), and the EABR thresholds in children with stenotic versus normal cochlear nerve canals. METHOD: Sixteen children with profound sensorineural hearing loss were included in this study: 8 with CNCs (CNCs group) and 8 with normal cochlear nerve canals (control group). All children underwent cochlear implantation with full insertion of all electrodes. EABR was performed 6 months postoperatively in both groups. RESULTS: The EABR extraction rate was 100% in children with normal cochlear nerve canals and only 50% in children with CNCs. EABR thresholds were significantly higher in children with CNCs of electrodes No. 11and 22 than in children with normal cochlear nerve canals (P < 0.05 for both comparisons). There was no significant difference in EABR thresholds among electrode No. 1, 11 and 22 in CNCs group (P > 0.05 for all comparisons); while in the control group, the EABR threshold at electrode No 22 was lower than those at both electrodes No. 11 and 1 (P < 0.05 for both comparisons), and the EABR threshold at electrode No. 11 was also lower than that at electrode No. 1 (P < 0.05). CONCLUSION: The EABR thresholds in children with normal cochlear nerve canals vary according to the different locations of electrodes in the cochlea; while in children with CNCs, there was no significant difference among different electrode locations. The EABR thresholds in CNCs children were higher than those of children with normal cochlear nerve canals at electrode 11 and 22.

LXRα Regulates Hepatic ChREBPα Activity and Lipogenesis upon Glucose, but Not Fructose Feeding in Mice.

Liver X receptors (LXRα/ß) and carbohydrate response element-binding proteins (ChREBPα/ß) are key players in the transcriptional control of hepatic de novo lipogenesis. LXRα/ß double knockout (LXRα-/-/ß-/-) mice have reduced feeding-induced nuclear O-linked N-acetylglucosamine (O-GlcNAc) signaling, ChREBPα activity, and lipogenic gene expression in livers, suggesting important roles for LXRs in linking hepatic glucose utilization to lipid synthesis. However, the role of LXRs in fructose-induced ChREBP activation and lipogenesis is currently unknown. In this study, we studied the effects of high fructose or high glucose feeding on hepatic carbohydrate metabolism and lipogenic gene expression in livers from fasted (24 h) and fasted-refed (12 h) wild type and LXRα knockout (LXRα-/-) mice. Hepatic lipogenic gene expression was reduced in glucose fed, but not fructose fed LXRα-/- mice. This was associated with lower expression of liver pyruvate-kinase (L-pk) and Chrebpß, indicating reduced ChREBPα activity in glucose fed, but not fructose fed mice. Interestingly, ChREBP binding to the L-pk promoter was increased in fructose fed LXRα-/- mice, concomitant with increased glucose-6-phosphatase (G6pc) expression and O-GlcNAc modified LXRß, suggesting a role for LXRß in regulating ChREBPα activity upon fructose feeding. In conclusion, we propose that LXRα is an important regulator of hepatic lipogenesis and ChREBPα activity upon glucose, but not fructose feeding in mice.

TCDD-inducible poly-ADP-ribose polymerase (TIPARP/PARP7) mono-ADP-ribosylates and co-activates liver X receptors.

Members of the poly-ADP-ribose polymerase (PARP) family catalyse the ADP-ribosylation of target proteins and are known to play important roles in many cellular processes, including DNA repair, differentiation and transcription. The majority of PARPs exhibit mono-ADP-ribosyltransferase activity rather than PARP activity; however, little is known about their biological activity. In the present study, we report that 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-inducible poly-ADP-ribose polymerase (TIPARP), mono-ADP-ribosylates and positively regulates liver X receptor α (LXRα) and LXRß activity. Overexpression of TIPARP enhanced LXR-reporter gene activity. TIPARP knockdown or deletion reduced LXR regulated target gene expression levels in HepG2 cells and in Tiparp(-/-)mouse embryonic fibroblasts (MEFs) respectively. Deletion and mutagenesis studies showed that TIPARP's zinc-finger and catalytic domains were required to enhance LXR activity. Protein interaction studies using TIPARP and LXRα/ß peptide arrays revealed that LXRs interacted with an N-terminal sequence (a.a. 209-236) of TIPARP, which also overlapped with a putative co-activator domain of TIPARP (a.a. 200-225). Immunofluorescence studies showed that TIPARP and LXRα or LXRß co-localized in the nucleus.In vitroribosylation assays provided evidence that TIPARP mono-ADP-ribosylated both LXRα and LXRß. Co-immunoprecipitation (co-IP) studies revealed that ADP-ribosylase macrodomain 1 (MACROD1), but not MACROD2, interacted with LXRs in a TIPARP-dependent manner. This was complemented by reporter gene studies showing that MACROD1, but not MACROD2, prevented the TIPARP-dependent increase in LXR activity. GW3965-dependent increases in hepatic Srebp1 mRNA and protein expression levels were reduced in Tiparp(-/-)mice compared with Tiparp(+/+)mice. Taken together, these data identify a new mechanism of LXR regulation that involves TIPARP, ADP-ribosylation and MACROD1.

Neural Response Telemetry Thresholds in Patients with Cochlear Nerve Canal Stenosis.

OBJECTIVE: To explore neural response telemetry (NRT) thresholds in patients with stenotic versus normal cochlear nerve canals. STUDY DESIGN: Case series with chart review. SETTING: Tertiary referral center. SUBJECTS AND METHODS: Thirty pediatric patients with profound sensorineural hearing loss in at least 1 ear and no benefit from amplification underwent computed tomography imaging of the temporal bones. They were divided into 3 groups according to the diameter of the cochlear nerve canal: group A, <1.5 mm; group B, 1.5 to 1.7 mm; group C, 1.8 to 2.1 mm. All patients underwent cochlear implantation with full insertion of all electrodes. NRT was performed both intraoperatively and 6 months postoperatively in all patients; thresholds of electrodes 1, 11, and 22 were compared. RESULTS: Per analysis of variance, intraoperative and 6-month postoperative NRT thresholds were both significantly different among groups A, B, and C at electrodes 1 and 22 but not at electrode 11. On intergroup analysis, group A showed statistically higher thresholds than those of groups B and C; however, no difference was found between groups B and C. CONCLUSION: Cochlear nerve canal stenosis, defined as a canal diameter <1.5 mm, is associated with significantly increased NRT thresholds, which may play a role in postimplant performance.

Inhibition of L-type Ca(2+) current by ginsenoside Rd in rat ventricular myocytes.

BACKGROUND: Ginsenoside Rd (GSRd), one of the most abundant ingredients of Panax ginseng, protects the heart via multiple mechanisms including the inhibition of Ca(2+) influx. We intended to explore the effects of GSRd on L-type Ca(2+) current (I Ca,L) and define the mechanism of the suppression of I Ca,L by GSRd. METHODS: Perforated-patch recording and whole-cell voltage clamp techniques were applied in isolated rat ventricular myocytes. RESULTS: (1) GSRd reduced I Ca,L peak amplitude in a concentration-dependent manner [half-maximal inhibitory concentration (IC50) = 32.4 ± 7.1 µmol/L] and up-shifted the current-voltage (I-V) curve. (2) GSRd (30 µmol/L) significantly changed the steady-state activation curve of I Ca,L (V 0.5: -19.12 ± 0.68 vs. -16.26 ± 0.38 mV; n = 5, p < 0.05) and slowed down the recovery of I Ca,L from inactivation [the time content (ζ) from 91 ms to 136 ms, n = 5, p < 0.01]. (3) A more significant inhibitive effect of GSRd (100 µmol/L) was identified in perforated-patch recording when compared with whole-cell recording [65.7 ± 3.2% (n = 10) vs. 31.4 ± 5.2% (n = 5), p < 0.01]. (4) Pertussis toxin (G i protein inhibitor) completely abolished the I Ca,L inhibition induced by GSRd. There was a significant difference in inhibition potency between the two cyclic adenosine monophosphate elevating agents (isoprenaline and forskolin) prestimulation [55 ± 7.8% (n = 5) vs. 17.2 ± 3.5% (n = 5), p < 0.01]. (5) 1H-[1,2,4]Oxadiazolo[4,3-a]-quinoxalin-1-one (a guanylate cyclase inhibitor) and N-acetyl-l-cysteine (a nitric oxide scavenger) partly recovered the I Ca,L inhibition induced by GSRd. (6) Phorbol-12-myristate-13-acetate (a protein kinase C activator) and GF109203X (a protein kinase C inhibitor) did not contribute to the inhibition of GSRd. CONCLUSION: These findings suggest that GSRd could inhibit I Ca,L through pertussis toxin-sensitive G protein (Gi) and a nitric oxide-cyclic guanosine monophosphate-dependent mechanism.

Serglycin is part of the secretory repertoire of LPS-activated monocytes.

Monocytes play multiple roles in the immune system, and are active in both acute and chronic diseases. Patients exposed to bacterial infections depend on monocytes in defense reactions, but excessive immune reactions may also cause morbidity through systemic inflammatory responses. Few studies have addressed the importance of proteoglycans, and in particular, the hematopoietic serglycin, in such monocyte immune reactions. Adherent primary monocytes were cultured in absence and presence of LPS. Media were analyzed by ELISA for detection of serglycin. Lysed cell fractions were used to determine the mRNA level of serglycin. Monocytes were also cultured on chamber slides to investigate if serglycin could be detected intracellularly by immunocytochemistry. Monocytes secreted serglycin, and LPS-stimulation increased the secretion. Secretion of inflammatory cytokines increased to a larger extent than serglycin. mRNA levels of serglycin were also increased, suggesting both increased expression and secretion. Immunocytochemistry revealed the presence of serglycin in intracellular vesicles, many destined for secretion. Serglycin containing vesicles increased in number and size when the cells were exposed to LPS. Intracellular vesicle localization and secretion of the proteoglycan serglycin is shown for the first time in primary human monocytes. Monocyte activation by LPS increased the expression and secretion of serglycin, suggesting roles for serglycin in inflammatory processes.