NIK promotes metabolic adaptation of glioblastoma cells to bioenergetic stress.

Cancers, including glioblastoma multiforme (GBM), undergo coordinated reprogramming of metabolic pathways that control glycolysis and oxidative phosphorylation (OXPHOS) to promote tumor growth in diverse tumor microenvironments. Adaptation to limited nutrient availability in the microenvironment is associated with remodeling of mitochondrial morphology and bioenergetic capacity. We recently demonstrated that NF-κB-inducing kinase (NIK) regulates mitochondrial morphology to promote GBM cell invasion. Here, we show that NIK is recruited to the outer membrane of dividing mitochondria with the master fission regulator, Dynamin-related protein1 (DRP1). Moreover, glucose deprivation-mediated metabolic shift to OXPHOS increases fission and mitochondrial localization of both NIK and DRP1. NIK deficiency results in decreased mitochondrial respiration, ATP production, and spare respiratory capacity (SRC), a critical measure of mitochondrial fitness. Although IκB kinase α and ß (IKKα/ß) and NIK are required for OXPHOS in high glucose media, only NIK is required to increase SRC under glucose deprivation. Consistent with an IKK-independent role for NIK in regulating metabolism, we show that NIK phosphorylates DRP1-S616 in vitro and in vivo. Notably, a constitutively active DRP1-S616E mutant rescues oxidative metabolism, invasiveness, and tumorigenic potential in NIK-/- cells without inducing IKK. Thus, we establish that NIK is critical for bioenergetic stress responses to promote GBM cell pathogenesis independently of IKK. Our data suggest that targeting NIK may be used to exploit metabolic vulnerabilities and improve therapeutic strategies for GBM.

Soybean oligosaccharides attenuate odour compounds in excreta by modulating the caecal microbiota in broilers.

Abatement of odour emissions in poultry production is important to ensure the quality and safety in the poultry industry as well as for the benefit of the environment. This study was conducted to compare the effects of supplementation with different amounts of dietary soybean oligosaccharides (SBO) and chlortetracycline (CHL) on the major odour-causing compounds in the excreta and on the caecal microbiota in broiler chickens. One-day-old broiler chickens were subjected to a 42-day experiment involving 6 treatments with 6 replicates per treatment (10 birds/cage). The treatments were as follows: negative control (NC; basal diet), positive control (PC; basal diet supplemented with CHL) and basal diet supplemented with 0.5, 2.0, 3.5 and 5.0 g/kg SBO (0.5SBO, 2.0SBO, 3.5SBO and 5.0SBO, respectively). Fresh excreta were sampled for the analysis of odour compounds by HPLC. Caecum content was collected for the caecal microbiota analysis through 16S rRNA sequencing. Results showed that on day 42, the excreta indole concentration in the broilers fed with 2.0SBO, 3.5SBO and 5.0SBO and PC was significantly decreased (P < 0.01) compared with that in the NC-fed broilers. The excreta skatole concentration (P < 0.001) and pH (P < 0.05) were also decreased by SBO and CHL, and they were lowest in birds fed with 3.5SBO. The formate concentrations in birds fed with 3.5SBO and 5.0SBO were higher than those in birds fed with the other diets (P < 0.001). Similarly, acetate concentration (P = 0.003) was increased in birds fed with 3.5SBO. Deep sequencing of 16S rRNA revealed that the composition of the caecal microbial digesta was slightly or significantly changed by the SBO or by the CHL supplementation, respectively. The SBO supplementation decreased the abundance of Bacteroides, Bilophilaand Escherichia, which were associated with the skatole and indole concentrations in the excreta. By contrast, the CHL supplementation demonstrated a strong tendency to enrich Ruminococcus and to reduce the abundance of Rikenella. These results indicated that supplementation with dietary SBO is beneficial in attenuating the concentration of odour-causing compounds and that it modulates the composition of caecal microbiota in broiler chickens.

Effect of typhaneoside on ventricular remodeling and regulation of PI3K/Akt/mTOR pathway.

BACKGROUND: This study aimed to investigate the effect of typhaneoside on ventricular remodeling and regulation of the PI3K/Akt/mTOR autophagy transduction pathway in rats with heart failure after myocardial infarction. METHODS: The effects of typhaneoside on the general condition of rats were observed in vivo using a rat model of heart failure after myocardial infarction had been established. The expression of serum N­terminal pro-brain natriuretic peptide (NT-proBNP), matrix lysin 2 (ST2), interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-α), matrix metalloproteinase 2 (MMP-2), and MMP-9 was detected via ELISA. A hypoxia/reoxygenation model was established to analyze the number and morphology of autophagosomes in vitro by transmission electron microscopy. Light chain 3 (LC3) variations were detected by immunofluorescence. Western blotting was used to assess LC3-II/LC3-I and p62 expression as well as p­Akt/Akt, p­mTOR/mTOR ratios. RESULTS: Compared with the sham group, the general condition scores of the rats in the model group decreased significantly, while the expression of serum NT-proBNP, ST2, IL-6, TNF-α, MMP-2, and MMP-9 increased. The number of autophagosomes in the drug-containing serum group was significantly reduced and the ratio of LC3-II/LC3-I was significantly decreased. The expression of P62 protein was increased, and the ratios of p­Akt/Akt and p­mTOR/mTOR were significantly increased. CONCLUSION: Typhaneoside regulates IL-6 and TNF-α as well as MMP-2 and MMP-9 in rats with heart failure after myocardial infarction. Typhaneoside can improve cardiac morphological structure and myocardial remodeling and enhance heart function. It may mediate autophagy inhibition in the cardiomyocyte anoxia/reoxygenation (A/R) pathway through the PI3K/Akt/mTOR autophagy transduction pathway.

Neoadjuvant therapy versus upfront surgery for borderline-resectable pancreatic cancer.

BACKGROUND: Neoadjuvant therapy is recommended for patients with borderline-resectable pancreatic cancer (BRPC). In this study, we compare survival outcomes of neoadjuvant therapy with upfront surgery. METHODS: From January 2011 to June 2016, 1415 patients underwent treatments for pancreatic cancer in Samsung Medical Center. Among them, 112 (7.9%) patients were categorized as BRPC by the NCCN 2016 guideline. They were classified by type of initial treatments into neoadjuvant group (NA, N.=26) and upfront surgery group (US, N.=86). RESULTS: The median survival duration of all patients was 18.3 months. Patients in the NA group had more T4 disease than those in the US group (38.5% in NA versus 15.1% in the US group; P=0.010). Arterial involvement was more frequent in the NA group (42.3% versus 15.1%; P=0.003). In the NA group, ten (38.5%) patients underwent surgery, and seven of them had complete R0 resection. In the US group, 83 (96.5%) patients received radical surgery, and 42 (48.8%) had R0 resection. In survival analysis according to intent to treat, the overall two-year survival rate was 51.1% in the US group and 36.7% in the NA group (P=0.001). However, among patients who underwent surgery (N.=96), the two-year overall survival rate was not significantly different between the two groups (P=0.089). According to involved vessels, the survival rate was not different between patients with arterial or both arterial and venous involvement and in patients with only venous involvement (P=0.649). CONCLUSIONS: It is necessary to demonstrate the efficacy of neoadjuvant therapy and to standardize the regimens through large-scale, multicenter, randomized controlled studies.

Oviposition Deterrents in Larval Frass of Potato Tuberworm Moth, Phthorimaea operculella (Lepidoptera: Gelechiidae).

The potato tuberworm moth (PTM) Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae) is one of the most damaging pests of potato Solanum tuberosum L. in warm temperate and subtropical areas. Our previous experiment showed that extracts of larval frass of PTM deterred oviposition of conspecific females. In this study, we investigated the identification of chemicals in larval frass that were influencing the oviposition of PTM by behavioral bioassays and electroantennography analysis in the laboratory. Frass was collected from third and fourth instar larvae and combined analysis of gas chromatography coupled with electroantennography (GC-EAD) of dichloromethane extracts showed that eight compounds from larval frass extracts elicited repeatable antennal responses from mated females. Seven EAD-active compounds in frass volatile extract were identified by gas chromatography-mass spectrometry (GC-MS) as linoleic acid, octadecanoic acid, tricosane, pentacosane, heptacosane, nonacosane, and cholesterol. Oviposition bioassays indicated that frass extracts had a deterrent effect on egg laying, the deterrent activity increased with the concentration of frass extracts, and the threshold value for statistical significance in oviposition deterrence was in the range of 20-200 mg frass per cage. Linoleic acid, pentacosane, heptacosane, nonacosane, and cholesterol in larval frass volatiles were found to play a key role in repelling oviposition in a dose-dependent manner. We suggest that the bioactive compounds in larval frass are responsible for repelling oviposition of PTM, and n-alkanes, especially pentacosane, strongly deter oviposition and may be considered as a potential oviposition deterrent for potential applications.

Oral immunisation with Taishan Pinus massoniana pollen polysaccharide adjuvant with recombinant Lactococcus lactis-expressing Proteus mirabilis ompA confers optimal protection in mice.

BACKGROUND: Proteus mirabilis poses a critical burden on the breeding industry, but no efficient vaccine is available for animals. METHOD: A recombinant Lactococcus lactis expressing the ompA of P. mirabilis was used to develop a vaccine. The mucosal and systemic immune responses of the recombinant vaccine were evaluated in mice after oral immunisation. The inhibition on P. mirabilis colonisation of vaccines was also determined. Moreover, Taishan Pinus massoniana pollen polysaccharides (TPPPS) were used as adjuvants to examine the immunomodulatory effects. RESULTS: The pure recombinant L. lactis vaccine significantly induced the production of specific IgA and IgG, IL-2, IL-4, IFN-γ, and T lymphocyte proliferation, and the immunised mice exhibited significant resistance to P. mirabilis colonisation. Notably, the TPPPS adjuvant vaccines induced higher levels of immune responses than the pure L. lactis. CONCLUSIONS: The L. lactis as a vaccine vehicle combined with TPPPS adjuvant provides a feasible method for preventing P. mirabilis infection.

The effect of botulinum neurotoxin type A on capsule formation around silicone implants: the in vivo and in vitro study.

This study confirms that botulinum neurotoxin type A (BoNT-A) decreases capsular contracture and elucidates a possible mechanism. Silicone blocks were implanted subcutaneously in 20 mice. The experimental groups received BoNT-A (1, 2·5 or 5 U) instilled into the subcutaneous pocket. After 30 days, periprosthetic capsules were harvested and evaluated. The effect of BoNT-A on the differentiation of human dermal fibroblasts to myofibroblasts in culture was examined by Western blot analysis. Changes in transforming growth factor-beta1 (TGF-ß1) expression in cultured fibroblasts were determined by enzyme-linked immunosorbent assay (ELISA). In in vivo study, the thickness of capsules (P < 0·05) and the number of alpha-smooth muscle actin (α-SMA)(+) cells in capsules (P < 0·05) were significantly decreased in the experimental groups. TGF-ß1 was significantly underexpressed in the experimental groups (P < 0·05). In in vitro study, BoNT-A did not significantly affect fibroblast viability. Western blot analysis showed that α-SMA protein levels were significantly decreased in the experimental groups (P < 0·05). Based on ELISA, the amount of TGF-ß1 was significantly decreased in the experimental groups (P < 0·05), especially cells treated with a high dose of BoNT-A (P < 0·001). This study confirms that BoNT-A prevents capsular formation around silicone implants, possibly by blocking TGF-ß1 signalling and interrupting the differentiation of fibroblasts to myofibroblasts.

Protective effects of Aloe vera-based diets in Eimeria maxima-infected broiler chickens.

Aloes have been widely used for a broad range of pharmacological activities, including parasitic problems. Avian coccidiosis is the most costly and wide-spread parasitic disease in the poultry industry, and has been mainly controlled by the use of chemotherapeutic agents. Due to the emergence of drug-resistant strains, alternative control strategies are needed. In this study, the protective effects of Aloe vera-based diets were assessed in broiler chickens following oral infection with Eimeria maxima. Chickens were fed a regular diet supplemented with ground Aloe vera throughout the duration of the experiment beginning 2 days prior to infection with 1 × 10(4) sporulated oocysts of E. maxima. No significant differences were found in body weight gain or loss between the Aloe vera-supplemented and unsupplemented groups with or without E. maxima infections. Fecal oocyst shedding decreased significantly (p < 0.05) in all of the treatment groups that were supplemented with Aloe vera as compared to the unsupplemented group. Furthermore, the Aloe vera-supplemented group showed significantly fewer intestinal lesions (p < 0.05) than the unsupplemented group following infection. The findings of this study suggest that Aloe vera could be used an alternative treatment for controlling avian coccidiosis.

A phase I/II study of neoadjuvant liposomal doxorubicin, paclitaxel, and hyperthermia in locally advanced breast cancer.

PURPOSE: The prognosis for locally advanced breast cancer (LABC) patients continues to be poor, with an estimated five-year survival of only 50-60%. Preclinical data demonstrates enhanced therapeutic efficacy with liposomal encapsulation of doxorubicin combined with hyperthermia (HT). Therefore this phase I/II study was designed to evaluate the safety and efficacy of a novel neoadjuvant combination treatment of paclitaxel, liposomal doxorubicin, and hyperthermia. MATERIALS AND METHODS: Eligible patients received four cycles of neoadjuvant liposomal doxorubicin (30-75 mg/m(2)), paclitaxel (100-175 mg/m(2)), and hyperthermia. They subsequently underwent either a modified radical mastectomy or lumpectomy with axillary node dissection followed by radiation therapy and then eight cycles of CMF (cyclophosphamide, methotrexate, 5-fluorouracil) chemotherapy. RESULTS: Forty-seven patients with stage IIB-III LABC were enrolled and 43 patients were evaluable. Fourteen patients (33%) had inflammatory breast cancer. Combined (partial + complete) clinical response rate was 72% and combined pathological response rate was 60%. Four patients achieved a pathologically complete response. Sixteen patients were eligible for breast-conserving surgery. The cumulative equivalent minutes (CEM 43) at T90 (tenth percentile of temperature distribution) was significantly greater for those with a pathological response. Four-year disease-free survival was 63% (95% CI, 46%-76%) and the four-year overall survival was 75% (95% CI, 58-86%). CONCLUSIONS: Neoadjuvant therapy using paclitaxel, liposomal doxorubicin and hyperthermia is a feasible and well tolerated treatment strategy in patients with LABC. The thermal dose parameter CEM 43 T90 was significantly correlated with attaining a pathological response.

Sustained improvement of spinal muscular atrophy mice treated with trichostatin A plus nutrition.

Early treatment with the histone deacetylase inhibitor, trichostatin A, plus nutritional support extended median survival of spinal muscular atrophy mice by 170%. Treated mice continued to gain weight, maintained stable motor function, and retained intact neuromuscular junctions long after trichostatin A was discontinued. In many cases, ultimate decline of mice appeared to result from vascular necrosis, raising the possibility that vascular dysfunction is part of the clinical spectrum of severe spinal muscular atrophy. Early spinal muscular atrophy disease detection and treatment initiation combined with aggressive ancillary care may be integral to the optimization of histone deacetylase inhibitor treatment in human patients.

The effect of capsicum plaster in pain after inguinal hernia repair in children.

BACKGROUND: Capsicum plaster at a classical Chinese acupoint is an alternative to acupuncture, which has been used as a supplemental therapy to opioid analgesics for pain control during the postoperative period. We investigated the postoperative analgesic efficacy of capsicum plaster at Zusanli (ST-36) points after pediatric hernia repair. METHODS: This double-blind, sham-controlled study was designed in 108 children, aged 4 month to 9 year, undergoing unilateral hernia repair, and was randomly assigned to three treatment regimens: group Zusanli (Z)=capsicum plaster at Zusanli acupoints and placebo tape on the shoulder as a nonacupoint, group Sham (S)=capsicum plaster on the shoulders and placebo tape at Zusanli acupoints, and group control (C)=placebo tape at Zusanli acupoints and on the shoulder. The postoperative pain scores and analgesic requirements during 24 h postoperatively were assessed. RESULTS: Total meperidine consumption was significantly lower in group Z (0.87+/-0.35 mg.kg-1) compared with group C (1.27+/-0.41 mg.kg-1) and S (1.22+/-0.45 mg.kg-1) (P<0.001). The pain scores on both the objective pain scale (OPS) and the Children Hospital of Ontario Pain Scale (CHEOPS), were significantly lower in group Z compared with the other groups at 6 and 24 h postoperatively, but not at the 10 min and 1 h postoperative time periods. CONCLUSIONS: Placement of capsicum plaster at the Zusanli points reduces pain and postoperative opioid consumption in children undergoing inguinal hernia repair, but not in the first six postoperative hours.

Correlation between Helicobacter pylori infection and vitamin C levels in whole blood, plasma, and gastric juice, and the pH of gastric juice in Korean children.

BACKGROUND: It is well known that chronic gastritis induced by Helicobacter pylori may be associated with hypochlorhydria and may also be accompanied by low levels of vitamin C in plasma and gastric juice in adults. This study investigates the relationship between H. pylori infection and vitamin C levels in the blood, plasma and gastric juice and the gastric juice pH of Korean children. METHODS: During a 5-year period, multiple gastric antral biopsies were taken from 452 children who underwent gastroduodenoscopy. The biopsy specimen was inoculated into phenol red buffered urea broth and incubated for 48 hours to detect color changes. The histopathologic findings were evaluated using the Sydney System. Concentrations of vitamin C in whole blood, plasma, and gastric juice aspirate were measured using the 2,4-dinitrophenylhydrazine method. RESULTS: Four hundred fifty-two patients (228 boys, 224 girls) aged 1 to 15 years were enrolled in this study. H. pylori was detected in 112 patients (24.8%) using histology, whereas it was found in 204 patients (45.1%) using the urease test. One hundred seven patients (23.7%) had active gastritis, and 421 patients (93.1%) had chronic gastritis. Vitamin C levels in whole blood, plasma, and gastric juice exhibited significant negative correlation with the age of patients, the histologic density of H. pylori, the degree of active and chronic gastritis, and the severity of H. pylori infection (based on urease positivity and histologic density of H. pylori). Gastric juice pH was correlated with the degree of chronic gastritis and was significantly higher in urease-positive patients. CONCLUSIONS: The data demonstrate that vitamin C levels in whole blood, plasma, and gastric juice and the gastric juice pH in Korean children are closely related to the severity of H. pylori infection and the histologic changes in the stomach. These data suggest that vitamin C may play a role in determining infection and progression, and vitamin C supplementation may be an important axis for the management of H. pylori infection in children.

[The episodes of spreading depression during focal cerebral ischemia in rats and the effects of electroacupuncture].

AIM: To investigate the effects of electroacupuncture on the episodes of spreading depression (SD) in Wistar rats during focal cerebral ischemia. METHODS: A model of the middle cerebral artery occlusion (MCAo) in Wistar rats was performed with the intraluminal filament occlusion. The episodes of SD in focal cerebral ischemia within 3 hours and the effects of electroacupuncture on "He gu" point (LI 4) were detected using neuroelectrophysiological method and neuropathological method. RESULTS: Electroacupuncture can reduce the episodes of SD during focal cerebral ischemia. CONCLUSIONS: Electroacupuncture can reduce the infarct volume of the focal cerebral ischemia, its mechanism may relate to reducing the episodes of SD during focal cerebral ischemia.

Sorption and desorption of Eu and Yb on alumina: mechanisms and effect of fulvic acid.

The effects of pH, ionic strength and FA (fulvic acid) on the sorption and desorption of Eu(III) and Yb(III) on alumina were respectively investigated by using batch technique and radiotracers 152 + 154Eu and 169Yb. The distribution coefficients for sorption and desorption of Eu on alumina at pH 4.4, 4.6 and 5.7 in 1 mol/l NaCl solutions as a function of solid phase concentration were determined in the presence or absence of FA. The effects of pH, FA and ionic strength on the distribution coefficients for sorption and desorption of Yb on alumina were determined in 0.01-2.0 mol/l NaNO3. It was found that pH and FA influenced the sorption of Eu(III) and Yb(III) on alumina greatly. A surface hydrolysis model can satisfactorily and qualitatively explain the observations on bare alumina. The competition among the complexations of surface free hydroxyl groups, soluble and sorbed fulvic acids can satisfactorily and qualitatively explain the observations on the coated alumina.

[Effect of anticancer polypeptide from Buthus Martensii venom on immune function in the H22-bearing mice].

OBJECTIVE: To observe the effect of anticancer polypeptide from Buthus Martensii Venom (APBMV) on Immune function in the H22-bearing mice. METHODS: The MTT colorimetric method, homolysin assay, lymphocyte transformation test, delayed hypersensitivity assay and WBC-count of peripheral blood were used in this study. RESULTS: APBMV could obviously augment NK activity, promote proliferation of lymphocytes induced by Con A, potentiate the response of DTH induced by DNCB, antagonize the decrease of WBC in peripheral blood induced by 5-Fu in the H22-bearing mice. CONCLUSION: APBMV can obviously increase immune function in the H22-bearing mice and antagonize hypoimmunity immunodeficiency or immunodeficiency induced by chemotherapy or the tumor.

Residual oil fly ash exposure enhances allergic sensitization to house dust mite.

Epidemiological studies have shown an association between elevated levels of particulate matter air pollution and increased morbidity and hospital visits in asthmatics. Residual oil fly ash (ROFA) is a primary combustion particle containing sulfate and metals such as vanadium, nickel, and iron. In this study the effect of ROFA on sensitization to house dust mite (HDM) was examined in a Brown Norway rat model of pulmonary allergy. Rats were instilled via the trachea with 200 or 1000 micrograms ROFA 3 days prior to local sensitization with 10 micrograms HDM and were challenged with 10 micrograms HDM 14 days later. Immunological endpoints were examined at 2, 7, and 14 days after sensitization and at 2 and 7 days after challenge (16 and 21 days post-sensitization, respectively). Antigen-specific immunoglobulin E and associated immediate bronchoconstriction responses to antigen challenge were increased in the ROFA-treated groups compared with the HDM control group. Lymphocyte proliferation to antigen was enhanced at Days 7 and 21 in the bronchial lymphocytes of ROFA-treated groups. Bronchoalveolar lavage fluid (BALF) eosinophil numbers and lactate dehydrogenase were significantly increased in the 1000 micrograms ROFA group at Days 2 and 16, BALF total proteins were elevated at Days 2 and 7 in both ROFA-treated groups, and BALF interleukin (IL)-10 was elevated in the 1000 micrograms ROFA group at Day 2. These results suggest that ROFA has an adjuvant effect on sensitization to HDM.

Enhanced allergic responses to house dust mite by oral exposure to carbaryl in rats.

Epidemiological studies have demonstrated an association between use of carbamate insecticides, including carbaryl, and increased incidence of allergic asthma in farmers. In this study the effect of oral carbaryl exposure on the development of allergic responses to house dust mite (HDM) was examined in female Brown Norway rats. Rats were gavaged for 2 weeks with 0, 2, 10, or 50 mg/kg/day of carbaryl. They were sensitized with a subcutaneous injection of HDM in aluminum hydroxide adjuvant 3 days after the beginning of carbaryl exposure and challenged with antigen via the trachea 1 day after the final carbaryl ingestion. Two days after challenge, antigen-specific cell proliferation in pulmonary lymph nodes was significantly higher in the 50 mg/kg group than in controls, while antigen-specific splenocyte proliferation was decreased in groups dosed with 2, 10, and 50 mg/kg carbaryl. Total protein and lymphocyte number in bronchoalveolar lavage (BAL) fluid were also increased in the 50 mg/kg group. By 7 days after challenge, immune-mediated pulmonary inflammation (eosinophils), antigen-specific immunoglobulin (Ig) E level in serum, and antigen-specific IgE and IgA levels in BAL fluid were significantly elevated in the 50 mg/kg group. No apparent change was observed for lactate dehydrogenase and eosinophil peroxidase in BAL fluid, while the number of BAL macrophages were decreased in groups dosed with 10 and 50 mg/kg carbaryl. The results suggest that carbaryl may cause systemic immune suppression, while enhancing pulmonary allergic responses to house dust mite antigen.

Cloning, sequence analysis, and distribution of rat metallocarboxypeptidase Z.

A cDNA encoding human carboxypeptidase Z (CPZ), a novel metallocarboxypeptidase, was recently cloned (Song and Fricker, J. Biol. Chem., 272, 1054, 1997). In the present study, a cDNA encoding the rat homolog of CPZ was identified. As with the human form, rat CPZ contains an N-terminal domain of 120 amino acids that has 20% to 30% amino acid identity with the "frizzled" domain found on proteins that interact with Wnt, a protein involved in tissue polarity in early embryogenesis. Sequence analysis showed rat and human CPZ to be highly conserved within the frizzled domain (77% amino acid identity), the carboxypeptidase domain (91%), and the C-terminal 28 residues (78%). The entire rat CPZ protein has high sequence similarity with human CPZ (81% amino acid identity), moderate sequence similarity to human carboxypeptidase N (45%), human carboxypeptidase E (41%), and human carboxypeptidase M (33%), and less sequence similarity with other metallocarboxypeptidases. Northern blot analysis showed rat CPZ mRNA to be abundant in the placenta, with low to moderate levels in the brain, lung, thymus, and kidney. The BRL3A rat liver cell line and the PC12 rat adrenal cell line express high levels of CPZ mRNA. In situ hybridization analysis indicated that CPZ is expressed only in specific cell types. For example, in the brain, CPZ mRNA is present in leptomeningeal cells, but not in the majority of other cell types. This distribution in leptomeningeal cells is shared by AEBP1, a recently reported member of the metallocarboxypeptidase gene family. However, the distribution of CPZ and AEBP1 differ in pituitary and thyroid. Taken together, these studies suggest that CPZ functions in a range of cell types.

[Study on effect of Erigeron injection in prevention and treatment of cerebral ischemic injury].

OBJECTIVE: To study the effect of Erigeron injection (EI) on cerebral ischemic injury. METHODS: Animal model of brain ischemic/reperfusion was established in male Wistar rats and the effect of EI on proteinkinase C (PKC) activity, intraneuronal free calcium and regional cortex blood flow (rCBF) were observed. RESULTS: EI could prevent the activation of PKC, calcium overload and decrease of RCBF induced by cerebral ischemia/reperfusion. CONCLUSION: EI could prevent neuronal ischemic injury by inhibiting activation of PKC.

Evaluation of isofagomine and its derivatives as potent glycosidase inhibitors.

A pseudo-aza-monosaccharide and several pseudo-aza-disaccharide compounds were constructed based on replacement of the anomeric carbon with a nitrogen and the ring oxygen with a carbon. The inhibition constants of these compounds toward five different glycosidases, alpha-glucosidase, beta-glucosidase, isomaltase, alpha-mannosidase, and glucoamylase, were obtained. Isofagomine, the pseudo-aza-monosaccharide, shows a broad spectrum of strong inhibition against glycosidases. It is the most potent inhibitor of beta-glucosidase from sweet almonds reported to date and also a strong inhibitor of glucoamylase, isomaltase, and alpha-glucosidase. Isofagomine inhibits beta-glucosidase, glucoamylase, and isomaltase more strongly than 1-deoxynojirimycin where the ring oxygen has been replaced with a nitrogen. The alpha-1,6- linked pseudo-disaccharide showed very strong inhibition toward glucoamylase, being nearly as potent an inhibitor as acarbose. Pseudo-disaccharides in which the anomeric nitrogen was methylated to favor formation of either the alpha or beta substrate linkage generally had weakened inhibition for the glycosidases studied most likely due to steric interference with the various active sites. These results indicate that the presence of a basic group at the anomeric center is important for carbohydrase inhibition. The presence of a charged carboxylate group near the anomeric carbon which interacts with the basic nitrogen is suggested for these enzymes, particularly for beta-glucosidase. The presence of a second alpha-linked glucosyl residue is also critical for strong inhibition of glucoamylase.