[Safety and efficacy of ultrafiltration on heart failure patients with reduced ejection fraction and diuretic resistance: results from a single-center randomized controlled trial].

Objective: To investigate the safety and efficacy of ultrafiltration on diuretic sensitivity in heart failure patients with reduced ejection fraction and diuretic resistance. Methods: This was a single-center randomized controlled trial. A total of 148 heart failure patients with reduced ejection fraction admitted to the Hospital of Traditional Chinese Medicine of Xinjiang Uygur Autonomous Region from June 2010 to June 2020 were enrolled in this study, and these patients were randomly divided (ratio 1:1) into the ultrafiltration group (n=74) and the control group (n=74). All patients were treated with diuretics, cardiotonic, vasodilator and other comprehensive drugs according to relevant guidelines. After grouping, the patients in the control group were treated with standard treatment plan, while patients in the ultrafiltration group were treated with ultrafiltration on top of standard therapy. Diuretic drugs were discontinued during ultrafiltration, and intravenously furosemide (40 mg) was given immediately and 24 hours after the end of ultrafiltration. Clinical data including gender, age, complicated diseases, New York Heart Association (NYHA) function classification, etc. were collected. Effectiveness indicators include urine volume (the first 12-hour and 24-hour urine volume and the second 24-hour urine volume after using diuretic), body weight and dyspnea severity score. Safety indicators include systolic blood pressure, serum creatinine, serum Na+ concentration, blood K+ concentration and the number of deaths before and after intervention. Results: Two patients in the control group died due to worsening heart failure after randomization and were excluded in this study, 146 patients were finally analyzed (72 patients in the control group and 74 patients in the ultrafiltration group). There were 93 males, and the age was (68.3±11.2) years. There was no significant difference between patients in the ultrafiltration group and the control group in gender, age, body weight, course of disease, dyspnea severity score, NYHA function classification Ⅲ/Ⅳ, the proportion of patients with severe edema of both lower limbs, the proportion of patients with complicated diseases, and basic medication (all P>0.05). After using diuretics, the urine volume of the first 12-hour and 24-hour and the second 24-hour were significantly higher in the ultrafiltration group than in the control group (all P<0.05). Body weight decreased significantly after ultrafiltration treatment as compared with that before intervention in the ultrafiltration group (P<0.05). Compared with the control group, the dyspnea severity score was significantly improved in the ultrafiltration group (P<0.05). There was no significant difference in systolic blood pressure, serum creatinine, serum Na+ concentration, blood K+ concentration of patients between ultrafiltration group and control group before and after intervention (all P>0.05). During the clinical diagnosis and treatment, 2 male patients in the control group died, and the cause of death was aggravation of basic diseases complicated with acute heart failure and cardiogenic shock. There was no death in the ultrafiltration group, and there were no obvious clinical adverse events during and after ultrafiltration. Conclusion: Ultrafiltration therapy is safe and can improve diuretic sensitivity in heart failure patients with reduced ejection fraction and diuretic resistance.

[The efficacy and safety of ultrafiltration for patients with heart failure: results from a single-center randomized controlled study].

Objective: To evaluate the efficacy and safety of ultrafiltration in patients with heart failure. Methods: One hundred and thirty four cases of patients with heart failure, who hospitalized in our hospital from June 2010 to June 2016 were enrolled in this study. Random serial number was generated using SPSS 22.0 software, patients were then randomly divided into control group and ultrafiltration group with the proportion of 1∶1 (67 cases in each group). Patients in the control group received standard therapy. Patients in the ultrafiltration group received ultrafiltration therapy for 8 hours. Curative effect was evaluated after 8 hours treatment in the control group and after 12 hours in the ultrafiltration group. Following parameters were compared between the two groups: body weight, dyspnea score and 6 minutes walking distance as well as blood pressure, heart rate, Na(+) , K(+) , Cl(-), pH, HCO(3)(-), Hb, PLT, Cr, BUN levels. Results: (1)Two patients died during run-in process and eventually 132 cases were chosen for final analysis (65 cases in control group and 67 cases in the ultrafiltration group). Gender, age, type of heart failure, dyspnea score, body weight at baseline were similar between the two groups. (2)Post therapy, patients' body weight decreased obviously, while dyspnea score and 6 minutes walking distance increased significantly in the ultrafiltration group compared to baseline(all P<0.05), and the improvement was significantly greater compared to control group(all P<0.05). (3)The safety index comparison of two groups: blood pressure, heart rate, Na(+) , K(+) , Cl(-), pH, HCO(3)(-), Hb, PLT, Cr, and BUN were similar between the two groups at baseline and post therapy. Conclusion: Ultrafiltration therapy is safe and effective to treat patients with heart failure.

Molecular evolutionary analysis of the high-affinity K+ transporter gene family in angiosperms.

The high-affinity K(+) transporter (HKT) family comprises a group of multifunctional cation transporters widely distributed in organisms ranging from Bacteria to Eukarya. In angiosperms, the HKT family consists primarily of nine types, whose evolutionary relationships are not fully understood. The available sequences from 31 plant species were used to perform a comprehensive evolutionary analysis, including an examination of selection pressure and estimating phylogenetic tree and gene duplication events. Our results show that a gene duplication in the HKT1;5/HKT1;4 cluster might have led to the divergence of the HKT1;5 and HKT1;4 subfamilies. Additionally, maximum likelihood analysis revealed that the HKT family has undergone a strong purifying selection. An analysis of the amino acids provided strong statistical evidence for a functional divergence between subfamilies 1 and 2. Our study was the first to provide evidence of this functional divergence between these two subfamilies. Analysis of co-evolution in HKT identified 25 co-evolved groups. These findings expanded our understanding of the evolutionary mechanisms driving functional diversification of HKT proteins.

Molecular cloning and analysis of a receptor-like promoter of Gbvdr3 gene in sea island cotton.

Verticillium wilt caused by soil borne fungus Verticillium dahliae could significantly reduce cotton yield. The Ve1 homologous gene Gbvdr3 is resistant to Verticillium wilt. In order to understand of the function of the promoter Gbvdr3 in Gossypium barbadense, the promoter region of the receptor-like gene Gbvdr3 was obtained by genome walking, and the cis-element in the promoter was identified using the PLACE software in this study. The sequence analysis showed that the promoter contained elements related to stress resistance and light regulation. The cloned promoter was fused to the GUS reporter gene and transformed into Arabidopsis. GUS expression was specifically detected in roots, flowers, and seeds, suggesting that the expression of Gbvdr3 is tissue-specific. Separation and characterization analysis of the promoter of Gbvdr3 provides a platform for further research and application of this gene. Thorough understanding of the function of the Gbvdr3 promoter is important for better understanding of Gbvdr3 function. These results indicated that the promoter of Gbvdr3 was a tissue-specific promoter.

[Clinical analysis of acute disseminated encephalomyelitis in 44 cases].

Objective: To investigate the clinical features in 44 patients with acute disseminated encephalomyelitis (ADEM). Methods: Consecutive ADEM patients admitted to Neurology Department of the Third Affiliated Hospital of Sun yat-sen University during August 2009 to July 2014 were enrolled.Clinical and laboratory data of the patients were reviewed and analyzed. Results: Forty-four patients with ADEM based on the 2012 criteria were recruited, including 23 male and 21 female; 9 children, 11 teenagers and 24 adults.There were 23 monophasic ADEM (23/44, 52%) and 21 multiphasic ADEM (21/44, 48%). Fourteen patients (31.8%) had definite incentive factors within 2 weeks preceding the disease onset.The commonest presenting symptoms were fever (20/44, 45%), mental disorder (18/44, 41%), disturbance of consciousness (17/44, 39%) and seizure (12/44, 27%). The average EDSS score was (4.3±1.3), and the average mRS score was (2.7±0.9). Abnormal autoimmune antibodies were detected in 10 patients.Two patients were positive for NMO-IgG, and three patients were positive for oligoclonal bands.On MRI scanning, small lesions were observed in 18 of 44 patients (18/44, 41%); large confluent white matter lesions in 10 patients (10/44, 23%); symmetric bithalamic involvement in 12 patients (12/44, 27%). Patients were mainly treated with intravenous corticosteroids (40/44, 90.9%) and immunoglobulin G ( 13/44, 29.5%) in acute phase.Regular follow-up performed in 29 patients (65.9%), and the average follow-up time was (4.2±2.3) year.A monophasic course was found in 10 patients, and multiphasic course in 19 patients.After (2.5±2.3) years, patients with multiphasic ADEM experienced their first clinical relapse, and the relapse frequency was (3.3±1.4). The average EDSS score was (3.9±2.2), and the mRS score was (2.2±1.3) in their latest relapse.In follow-up MRI for (5.3±1.9) years, lesions in 18 patients (62.1%) were partially ameliorated, while 6 patients (20.7%) persisted, and new lesions appeared in 5 patients (17.2%). For the 13 multiphasic patients with regular treatment, intravenous corticosteroids (13/13, 100.0%) and immunoglobulin G (7/13, 53.8%) were still important treatments in the acute phase, while oral steroids (12/13, 92.3%) plus immunosuppressants including azathioprine, tacrolimus, cyclosporine and rituximab were chosen in the remission phase. Conclusions: ADEM is not uncommon in adults, presenting with multiphasic course, encephalopathy features and disseminated lesions on MRI.As it shows very heterogeneous characteristics, ADEM is best viewed as a "syndrome" rather than a specific disorder.

Isolation and identification of phase I metabolites of resibufogenin in rats.

1. Resibufogenin (1), a major bufadienolide of Chinese medicine Chan Su, had a wide range of pharmacological activities. In present work, the metabolism of 1 in male Sprague-Dawley rats was investigated by identifying the metabolites of resibufogenin excreted in rat bile. 2. Following an oral dose of 60 mg/kg resibufagenin, nine metabolites were isolated from bile of rats, and their structures were identified as 3-keto- resibufogenin (2), 3-epi-resibufogenin (3), 5ß-hydroxy-3-epi-resibufogenin (4), 1α, 5ß-dihydroxy-3-epi-resibufogenin (5), 3α, 5ß, 14α, 15ß-tetrahydroxyl-bufa- 20, 22-dienolide (6), 3α, 14α, 15ß-trihydroxy-bufa-20, 22-dienolide (7), 3-epi- 5ß-hydroxy-bufalin (8), 12α, 16ß-dihydroxy-3-epi-resibufogenin (9), and 5ß, 16ß-dihydroxy-3-epi-resibufogenin (10), respectively, on the basis of widely spectroscopic methods including 2D-NMR technology. It is first time to describe the metabolites of 1 in vivo, and metabolites 5-7 and 9-10 are novel. 3. On the basis of these identified metabolites, a possible metabolism pathway for 1 in rats has been proposed. This is the first systematic study on the phase I metabolites of resibufogenin.

Metabolic changes of human dental pulp after rapid palatal expansion.

OBJECTIVES: To investigate rapid palatal expansion (RPE)-induced metabolic changes in human dental pulp by measuring the expression and activity of aspartate aminotransferase (AST). METHODS: mRNA and protein levels of AST in human dental pulp were measured by quantitative real-time polymerase chain reaction and Western blot, respectively. Furthermore, the activity of AST was measured by a full automatic biochemical analyzer. RESULTS: AST mRNA and protein levels were found to be expressed in normal dental pulp. Moreover, the expression of AST was increased significantly after 14 days of RPE and then decreased at 1 month in retention. Three and 6 months after RPE, the AST expression level was gradually decreased to its baseline level. Similarly, AST activity was significantly elevated after 14 days of RPE, which was then down-regulated at 1 month in retention but was still kept at a higher level as compared with the control group. The enzymatic activity of AST was slowly decreased to its baseline level at 3 and 6 months in retention. CONCLUSIONS: These results showed that significant reversible metabolic changes occurred in dental pulp during RPE, which revealed the high capacity of the pulp tissue for adaptation to this orthopedic method.

Isolation and identification of metabolites of osthole in rats.

Osthole (Ost), one of the major components of Cnidium monnieri (L.) Cusson, is had the structure of an isopentenoxy-coumarin with a range of pharmacological activities. In the present study, the metabolism of Ost in male Sprague-Dawley rats was investigated by identifying Ost metabolites excreted in rat urine. Following an oral dose of 40 mg/kg Ost, 10 phase I and 3 phase II metabolites were isolated from the urine of rats, and their structures identified on the basis of a range of spectroscopic data, including 2D-NMR techniques. These metabolites were fully characterized as 5'-hydroxyl-osthole (M-1), osthenol (M-2), 4'-hydroxyl-osthole (M-3), 3, 5'-dihydroxyl-osthole (M-4), 5'-hydroxyl-osthenol (M-5), 4'-hydroxyl-2', 3'-dihydro-osthenol (M-6), 4'-hydroxyl-osthenol (M-7), 3, 4'-dihydroxyl-osthole (M-8), 2', 3'-dihydroxyl-osthole (M-9), 5'-hydroxyl-2', 3'-dihydroosthole (M-10), osthenol-7-O-ß-D-glucuronide (M-11), osthole-4'-O-ß-D-glucuronide (M-12) and osthole-5'-O-ß-D-glycuronate (M-13). This is the first identification of M-1, M-3 to M-13 in vivo. On the basis of the metabolites profile, a possible metabolic pathway for Ost metabolism in rats has been proposed. This is the first systematic study on the phases I and II metabolites of 8-isopentenoxy-coumarin derivative.

Identification of cinobufagin metabolites in the bile of rats.

Cinobufagin (1) is a major bufadienolide in ChanSu (a traditional Chinese medicine) with a wide range of pharmacological activities. In this paper, the in vivo metabolites of 1 in rats were studied. Nine metabolites were isolated from the bile of rats, and their structures were identified as: desacetylcinobufagin (2), 3-ketodesacetylcinobufagin (3), 3-epi-desacetylcinobufagin (4); 5beta-hydroxy-3-epi-desacetylcinobufagin (5), 1alpha-hydroxy-3-epi-desacetylcinobufagin (6), 12beta-hydroxy-3-epi-desacetylcinobufagin (7), 1beta-hydroxy-3-epi-desacetylcinobufagin (8), 1alpha,5alpha-dihydroxy-3-epi-desacetylcino-bufagin (9), and 2alpha, 5beta-dihydroxy-3-epi-desacetylcinobufagin (10), respectively, on the basis of widely spectroscopic studies including two-dimensional-nuclear magnetic resonance (NMR). Among them, metabolites 6-10 are new compounds. The results show that hydroxylation is the main reaction involved in metabolism of 1, and the preferred hydroxylation sites were C-1 and C-5.

[Observation on clinical efficacy and prognosis analysis of endoscopic surgery treatment for rT1-rT2 recurrent nasopharyngeal carcinoma].

Objective:To investigate the feasibility of endoscopic surgery for treatment of rT1-rT2 recurrent nasopharyngeal carcinoma.Method: The clinical data and of 57 patients who had recurrence of the primary lesion after treatment of nasopharyngeal carcinoma from February 2011 to December 2015 were retrospectively analyzed. The patients were re-staged according to Union for International Cancer Control(UICC, 2010) staging system for nasopharyngeal carcinoma before surgery. Patients suitable for surgery underwent endoscopic surgery to remove nasopharyngeal lesions; those combined with cervical lymph node metastases underwent cervical lymph node dissection at the same time; patients with positive surgical margins of pharyngeal lesions and cervical lymph node extramembranous filtration were treated with radiotherapy combined with chemotherapy; patients unsuitable for surgery were treated with radiotherapy combined with chemotherapy directly. All patients were followed up regularly to observe clinical efficacy and survival. Result:Fifty-seven patients were re-staged according to UICC(2010) staging system for nasopharyngeal carcinoma: 19 cases in stage Ⅰ,30 cases in stage Ⅱ, 6 cases in stage Ⅲ and 2 cases in stage Ⅳ,including 27 cases in stage rT1, 30 cases in stage rT2, and 43 cases in stage rN0,6 cases in stage rN1,6 cases in stage rN2,2 cases in stage rN3. Forty-four cases of primary lesions were sected for endoscopic surgery. Patients combined with cervical lymph node metastases underwent cervical lymph node dissection at the same time, with 6 cases of positive surgical margins of pharyngeal lesions and 4 cases of cervical lymph node extramembranous infiltration, who were treated with radiotherapy combined with chemotherapy after surgery. Thirteen patients received radiotherapy combined with chemotherapy directly. At a median follow-up of 36 months, the 3-year overall survival rate of 57 patients was 61.4%. The 3-year overall survival rates of patients in stage Ⅰ, Ⅱ, Ⅲ and Ⅳ were 73.7%, 63.3%, 33.3%, 0.0% respectively. Kaplan-Meier survival curve analysis showed a significant difference in the overall survival rate of patients in different stages(P=0.002). The 3-year overall survival rates of rT1 and rT2 patients were 63.0%, 60.0% respectively, and KaplanMeier survival curve analysis showed no significant difference in the overall survival rate between rT1 and rT2 patients(P=0.707). The 3-year overall survival rates of patients in stages rN0, rN1, rN2, rN3 were 69.8%, 50.0%, 33.3%, 0.0% respectively, and Kaplan-Meier Survival curve analysis showed a significant difference in overall survival between patients in different rN stages(P=0.002). The 3-year overall survival rate was 68.2% in 44 surgical patients, and 38.5% in 13 non-surgical patients. Kaplan-Meier survival curve analysis showed significant difference in overall survival rate between surgical and non-surgical patients(P=0.014).Conclusion: Endoscopic surgery for recurrent nasopharyngeal carcinoma is a safe and effective treatment to improve survival.

Influences of miR-155/NF-κB signaling pathway on inflammatory factors in ARDS in neonatal pigs.

OBJECTIVE: Acute respiratory distress syndrome (ARDS) is greatly threatening human health with high morbidity and mortality. The pathogenesis of ARDS is closely related to the inflammatory response in patients. The micro-ribonucleic acid (miR)-155/nuclear factor-κB (NF-κB) signaling pathway is crucial in regulating the expression of inflammation-related genes. Therefore, the influences of miR-155/NF-κB signaling pathway on inflammatory factors in ARDS in neonatal pigs were explored in this study. MATERIALS AND METHODS: The model of ARDS in neonatal pigs was established first. The expression levels of miR-155, NF-κB-related proteins, and inflammatory factors in model group and control group were detected, and their differences were compared. Moreover, after treatment with the miR-155/NF-κB signaling pathway inhibitor, the changes of the inflammatory factors expression in ARDS neonatal pigs were observed at different time points. RESULTS: In the model group, the levels of miR-155 and NF-κB-related proteins were significantly increased, and the levels of inflammatory factors, including interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNF-α), and IL-6, were also increased synchronously. However, the levels of IL-4 and IL-10 declined significantly. In addition, it was proved that after treatment with the inhibitor in model group the mRNA expressions of miR-155/NF-κB signaling pathway-related proteins were significantly inhibited, and the levels of IL-1ß, TNF-α, and IL-6 were also significantly inhibited (p<0.05). The levels of IL-4 and IL-10 remarkably rose after treatment with the inhibitor for 24 h (p<0.05). CONCLUSIONS: The miR-155/NF-κB signaling pathway influenced the changes of inflammatory factors in ARDS in neonatal pigs, which might be a potential target for eliminating the inflammatory response after ARDS in neonatal pigs.

[Diagnostic value of narrow-band imaging in detection of recurrent nasopharyngeal carcinoma].

Objective: To evaluate the diagnostic value and feasibility of narrow-band imaging in detection of recurrent nasopharyngeal carcinoma (NPC). Methods: One thousand three hundred and sixty-four NPC patients who had completed NPC treatment were enrolled. All patients were followed-up with imaging, serological examination of EB virus and nasopharyngeal endoscopy(WL and NBI mode), in which (1) both white light (WL) and NBI modes were done; (2) positive endoscopic patients were given nasopharyngeal biopsy; (3) using histologic finding as criterion standard, the sensitivity, specificity, accuracy and Yonden's index of two modes were compared. Kappa index was used to evaluate the consistency between the two modes and pathological results respectively; (4) the positive rates of WL and NBI in patients with early recurrent (stage Ⅰ+ Ⅱ) were compared. Results: A total of 265 cases were suspected as having recurrent lesions by endoscopy in WL mode and 68 cases of them were pathologically diagnosed as having NPC; and 82 cases were suspected as having recurrent lesions by endoscopy in NBI mode and 74 cases of them were pathologically diagnosed as having NPC. The sensitivity, specificity, accuracy and Yonden's index of WL mode were 91.89%, 0, 25.09% and -0.0811, respectively, with a kappa of -0.045; the sensitivity, specificity, accuracy and Yonden's index of NBI mode were 100.00%, 95.94%, 97.05% and 0.9594, respectively. Conclusion: NBI has higher sensitivity, specificity, early diagnosis rate and Yonden's index than WL.

Temporomandibular joint ankylosis contributing to coronoid process hyperplasia.

The purpose of this study was to investigate the association between coronoid process hyperplasia and temporomandibular joint (TMJ) ankylosis and to analyze the pathological mechanism and clinical significance of coronoid process hyperplasia. Forty-four patients treated for TMJ ankylosis between January 2007 and December 2014 were studied retrospectively; 176 patients with normal TMJs served as controls. The original DICOM data were used to reconstruct the jaw, and a three-dimensional cephalometric analysis (SimPlant Pro software version 11.04) was performed to assess the association between the severity of TMJ ankylosis and the height of the coronoid process. The height of the coronoid process was 20.41±5.00mm in the case group and 14.86±2.67mm in the control group; there was a significant difference between the two groups (P<0.001). Long-standing TMJ ankylosis contributes to coronoid process hyperplasia. Therefore, attention should be drawn to the coronoid process in patients with TMJ ankylosis. A coronoidectomy together with arthroplasty is recommended in patients with TMJ ankylosis.

Reactivity patterns for redox reactions of monomer forms of myoglobin, hemocyanin and hemerythrin.

Electron-transfer reactions of myoglobin, hemocyanin and hemerythrin with the inorganic complexes [Fe(CN)6]3- (oxidant) and [Co(sep)]2+ (reductant) are considered. Rate constants kFe (25 degrees C) have been determined for the [Fe(CN)6]3- (410 mV) oxidation of horse deoxyMb, I = 0.100 M (NaCl). From the decrease in kFe over the range pH 5.5 to 9.0 a pKa of less than 6.2 is obtained, most likely due to the involvement of the heme propionate(s). At the higher pH values the rate constant is 1.2 x 10(6) M-1 s-1. Rate constants kCo (25 degrees C) for the [Co(sep)]2+ (-260 mV) reduction of metMb are also pH-dependent, pKa = 8.82, corresponding to acid dissociation of the H2O axially coordinated to the Fe(III). The rate constant for the aqua-met form is 2.8 x 10(3) M-1 s-1 at pH values less than 7.0. In contrast, no reaction is observed for the deoxy and met forms of P. interruptus hemocyanin monomer subunit a with the same two complexes (k less than 10(2) M-1 s-1). Comparisons are made with rate constants for hemerythrin, also as the monomer, which have been determined previously. Rate constants for the reactions of deoxy forms with the neutral small molecules, here O2 and H2O2, are also considered. Whereas the reactions of [Fe(CN)6]3- and [Co(sep)]2+ are at the protein surface, those of O2 and H2O2 are at the active site. Hemocyanin with the more buried (approximately 20 A) active site compared with myoglobin (3.8 A) and hemerythrin (6.3 A), does not readily undergo electron transfer with reagents at the surface. However, with the small molecules O2 and H2O2 penetration of the surrounding peptide occurs, with reaction at the active site. Rate constants for the three proteins are now of similar magnitude, and in the range (2.3-7.8) x 10(7) M-1 s-1 for O2, and 10.9 to 3600 M-1 s-1 for H2O2.

Chronic relapsing experimental autoimmune encephalomyelitis: effects of insulin-like growth factor-I treatment on clinical deficits, lesion severity, glial responses, and blood brain barrier defects.

Chronic relapsing experimental autoimmune encephalomyelitis (crEAE), a model for multiple sclerosis, was used to test 2 regimens of insulin-like growth factor-I (IGF-I) treatment. We induced crEAE by injecting 3x10(7) myelin basic protein-(MBP) sensitized lymph node cells into adult female SJL/J mice. Fifty-one mice, divided randomly into 4 groups, were used in the first trial. Two groups received IGF-I (a gift of Cephalon, Inc.) 0.6 mg/kg/d subcutaneously from day 7 to day 16 and the other two groups received placebo injections. IGF-I treatment reduced clinical deficits during the first attack and during 2 subsequent relapses. Image analysis of immunostained and histological sections showed that IGF-I treatment reduced BBB defects and both the numbers and sizes of inflammatory, demyelinating, and demyelinated lesions. Twelve mice that had recovered from their first attack were used in our second trial to evaluate possible adverse effects of prolonged treatment with a higher dose of IGF-I. Six received 1.2 mg/kg/d for 6 weeks (days 19-63). No adverse effects of IGF-I treatment were identified. The eyes, hearts, livers, and kidneys of IGF-I-treated mice were normal histologically and their spleens also appeared normal except for mild to moderate microscopic increases in lymphopoesis. Our results suggest that prolonged IGF-I treatment is well tolerated and that the anti-inflammatory effects of IGF-I have a major role in reducing clinical deficits and lesion severity in crEAE. These effects, if present in multiple sclerosis, may benefit patients with this disease.

Targeting of green fluorescent protein to secretory granules in oxytocin magnocellular neurons and its secretion from neurohypophysial nerve terminals in transgenic mice.

Oxytocin (OT) is a hypothalamic nonapeptide that is synthesized as part of a larger precursor protein that also contains an approximately 10-kDa protein called neurophysin at its C-terminus. This precursor protein is trafficked through the regulated secretory pathway into secretory granules and then axonally transported to and secreted from nerve terminals in the neural lobe of the pituitary. In this paper, we show that the AI-03 transgene that contains enhanced green fluorescent protein (EGFP) fused to the end of the neurophysin at the C-terminus of the OT pre-prohormone, is expressed selectively in OT-magnocellular neurons and is trafficked to secretory granules in transgenic mice. The EGFP-containing secretory granules are then transported to OT-neurosecretory terminals in the neurohypophysis, where the EGFP fluorescence undergoes depolarization-induced calcium-dependent secretion. The endogenous fluorescence in the neural lobes is sufficiently intense to image secretory events in individual OT nerve terminals (neurosecretosomes) isolated from the posterior pituitaries in these transgenic mice.

Oxidation of residue 45 mutant forms of pig deoxymyoglobin with [Fe(CN)6]3-.

The effect of replacement of the highly conserved Lys45 residue in pig myoglobin (Mb) with His, Ser, Glu, and Arg has been investigated. Rate constants/M-1 s-1 at 25 degrees C and pH 8.0, I = 0.100 M (NaCl), for the oxidation of deoxyMb with [Fe(CN)6]3- have been determined, and are for wild-type Lys45 (2.83 x 10(6)), His45 (1.02 x 10(6)), Ser45 (1.12 x 10(6)), Glu45 (0.87 x 10(6)), and Arg45 (3.06 x 10(6)). It is concluded that charge on the residue at position 45 has only a mild effect on reactivity, and that this is unlikely to be the site for electron transfer.

Biosynthesis in vitro of core lacto-series glycosphingolipids by N-acetyl-D-glucosaminyltransferases from human colon carcinoma cells, Colo 205.

Two N-acetyl-D-glucosaminyltransferases have been detected in human colon carcinoma Colo 205 cells. These enzymes catalyze the biosynthesis in vitro of the core-glycolipid of Type 1 and Type 2 lacto-series antigens and of the polylactosamine-containing longer chain antigenic structures, respectively. The first enzyme, GlcNAcT-1, which catalyzes the formation of lactotriosylceramide [LcOse3Cer, beta-D-GlcpNAc-(1----3)-LcOse2Cer, the core for all lacto-series Type 1 and Type 2 chains] from lactosylceramide [beta-D-Galp-(1----4)-D-Glcp-Cer, LcOse2Cer] and UDP-GlcNAc shows optimum activity in the presence of nonionic detergent Triton CF-54. The other enzyme, GlcNAcT-2, which catalyzes the biosynthesis in vitro of iLcOse5Cer [beta-D-GlcpNAc-(1----3)-nLcOse4Cer, the core for polylactosamine-containing antigens] from nLcOse4Cer [beta-D-Galp-(1----4)-LcOse3Cer] and UDP-GlcNAc, is optimally active with the zwitterionic detergent, Zwittergent 3-14, when membrane-bound. Both of these activities, however, can be extracted from the membrane by use of a nonionic detergent. Triton X-114, with nearly the same efficiency. These two transferases showed different pH optima, different cation and anion effects, and differential heat-inactivation patterns at 55 degrees. Permethylation studies of the radioactive products isolated from both of the enzyme-catalyzed reactions using respective 3H-substrates and nonradioactive UDP-GlcNAc showed the presence of 2,4,6-tri-O-methylgalactose in the hydrolyzed products. This indicated the presence of a (1----3)-linked beta-D-GlcpNAc group at the nonreducing end in both cases. The linkage of the beta-D-GlcpNAc group to the subterminal D-Gal residue in the two products was confirmed by an almost 90% cleavage of the terminal [3H]GlcNAc group by purified clam and papaya beta-D-hexosaminidases.

[Observation on hypotensive action of adrenomedullin and its distribution in tissues].

The effect of adrenomedullin (AM) on blood pressure and tension of aortic rings were studied in normal and in either spontaneous or renal hypertensive rats (SHR/RHR). The distribution characteristics of AM in viscera was also investigated. It was found that the hypotensive action of AM was far more prominent in hypertensive than in normotensive rats. The tension lowering action on the aortic rings is dose dependent and requires the presence of endothelium. Immunohistochemistry reactivity of AM of different degrees were also found in lung, kidney and myocardium, in addition to cells of pheochromocytoma and adrenomedulla. It was suggested that AM may play an important role in the regulation of blood pressure and visceral blood circulation.