[Exploration of phase angle used to construct PG-SGA nutritional assessment and prediction model for malignant tumor patients].

Objective: To explore the value of phase angle (PA) in constructing a predictive model of nutrition evaluation for tumor patients. Methods: A retrospective analysis was performed on 1 129 patients with malignant tumors hospitalized in the Cancer Center of Changzhi People's Hospital from June 2020 to February 2021. PA values of six parts of the body were measured by the body composition analyzer, including: left arm (LA), right arm (RA), left leg (LL), right leg (RL), the trunk (TR), and the whole body (WB). Patients' body mass index (BMI) was calculated and patient-generated subjective global assessment (PG-SGA) was assessed. The differences of PA values of six parts were compared and their correlations with BMI and PG-SGA in combination with age, gender and tumor disease types were analyzed, binary classification regression on BMI and PG-SGA was performed, and the functions of the best prediction model was fitted. Decision tree, random forest, Akaike information criterion in a Stepwise Algorithm (stepAIC) and generalized likelihood ratio test were used to select appropriate variables, and the logit logistic regression model was used to fit the data. Results: Comparing the PA values of six parts in pairs, it was found that the PA values of LA and RA, LL and RL, and TR and WB were linearly correlated and the coefficient was close to 1 (P<0.001). Binary classification regression was performed for BMI and PG-SGA, respectively. In order to make the data have clinical significance, 18.5 kg/m(2) was used as the classification point for BMI, 4 and 9 were used as the classification points for PG-SGA score, and the models of A, B and C were obtained. Suitable variables including PA-LA, PA-TR and tumor disease types were used as variables to fit BMI classification; BMI, PA-LA and age were used as variables to fit the PG-SGA model with 9 as the classification point. PA-LA, PA-TR, BMI, age and tumor disease types were used as variables to fit the PG-SGA model with 4 as the classification point. In this study, the predicted values of models A, B and C obtained by R-studio were imported into SPSS 26.0 software, and the cut-off values of classification were obtained by the receiver operating characteristic (ROC) curve. The ROC analytic results showed that the best cut-off values of Model A, B and C were 0.155, 0.793 and 0.295. Model A recommended when the probability is >0.155, a patient's nutritiond tatus should be classified as BMI < 18.5 kg/m(2) group. Model B recommended that PG-SGA<9 group be classified as the probability is >0.793. Model C recommended that PG-SGA < 4 group should be classified when probability is >0.295. Conclusions: The PG-SGA classification prediction model is simple to operate, and the nutritional status of patients can be roughly divided into three groups: normal or suspected malnutrition group (PG-SGA<4), moderate malnutrition group (4≤PG-SGA<9), and severe malnutrition group (PG-SGA≥9). This model can more efficiently predict the nutritional status of cancer patients, greatly simplify the nutritional assessment process, and better guide the standardized treatment of clinical malnutrition.

[Hepatic cystic echinococcosis complicated with tuberculous empyema misdiagnosed as hepatic and pulmonary cystic echinococcosis: one case report].

Hepatic cystic echinococcosis is a chronic parasitic disease caused by the infection with the larvae of Echinococcus granulosus in human or animal liver tissues. As a chronic active infectious disease, tuberculous empyema mainly invades the pleural space and then causes visceral and parietal pleura thickening. It is rare to present comorbidity for hepatic cystic echinococcosis and tuberculous empyema. This case report presents a case of hepatic cystic echinococcosis complicated with tuberculous empyema misdiagnosed as hepatic and pulmonary cystic echinococcosis, aiming to improve clinicians' ability to distinguish this disorder.

Preparation and physical properties of a Cr3Al film with a DO3 structure.

A Cr3Al compound with a DO3 structure has previously been predicted to be nearly half metal and a promising spintronics material; however, its synthesis has not been reported. Here, a Cr3Al compound with a DO3 structure is successfully prepared in thin-film form by the magnetron sputtering method. It was found that the substrate temperature is crucial to the atomic ordering, thin-film density and lattice constant. The lattice constant varies with different substrate temperatures and is smaller than the theoretical equilibrium lattice constant. Theoretical investigations on the electronic structures and magnetic properties indicate that the Cr3Al compound with a DO3 structure is a rare material with zero-gap half-metallic characteristics under an experimental lattice constant of 5.83 Å. The experimental result is in agreement with the theoretical results in magnetization, and the Cr3Al compound synthesized in this work exhibits semi-metallic-like electrical transport characteristics and positive magnetoresistance of greater than 2% in the temperature range 2-250 K.

[Effects of perioperative transfusion of blood components on the long-term prognosis of hepatocellular carcinoma].

Objective: To investigate the effects of perioperative transfusion of blood components on the long-term prognosis of hepatocellular carcinoma (HCC) patients. Methods: A total of 339 patients with primary HCC who underwent curative hepatectomy between January 2003 and December 2010 at the Cancer Hospital, Chinese Academy of Medical Sciences were enrolled. The clinical data of the patients were retrospectively analyzed. These patients were divided into non-transfusion, fresh frozen plasma (FFP) transfusion only and concentrated red cells (CRC) transfusion groups. Disease-free survival and overall survival were estimated using the Kaplan-Meier method, and Cox regression was performed to identify clinicopathological factors related with survival. Results: Among the 339 patients, the 1-, 3- and 5-year disease-free survival rates were 63.1%, 35.4% and 22.4%, respectively, and the median disease-free survival was 22 months. While the 1-, 3- and 5-year overall survival rates were 90.5%, 69.5% and 56.4%, respectively, and the median overall survival was 72 months. The median disease-free survivals of the non-transfusion (n=181), FFP transfusion only (n=48) and CRC transfusion (n=110) groups were 28, 22 and 12 months, respectively, while the median overall survivals of the three groups were 99, 63 and 40 months respectively. Significant differences in the disease-free and overall survivals were observed among the three groups (both P<0.01). Multivariate Cox regression analyses showed that FFP transfusion only (HR=1.658, P=0.026), CRC transfusion (HR=1.470, P=0.030), serum alpha-fetoprotein>400 µg/L (HR=1.686, P=0.002), albumin<35 g/L (HR=1.782, P=0.047), tumor capsule (HR=0.597, P=0.012), tumor necrosis (HR=1.820, P=0.001) and the TNM stage Ⅲ or above (HR=2.537, P=0.000) were independent predictors of overall survival after hepatectomy. Conclusion: Both perioperative FFP only transfusion and CRC transfusion may have detrimental effect on the long-term prognosis of HCC.

Protective effect of ischaemic preconditioning in total knee arthroplasty.

OBJECTIVE: To investigate the genomic response induced by ischaemic preconditioning (IPC) in muscle biopsies taken from the operative leg of total knee arthroplasty patients. MATERIALS AND METHODS: The gene expression profile GSE21164 was extracted from Gene Expression Omnibus (GEO) database. Patients undergoing primary knee arthroplasty were randomized to control and treatment (IPC) groups. Muscle biopsies were taken from the quadriceps muscle of the operative knee at the immediate onset of surgery (T0) and at 1 hour into surgery (T1). Limma package of R language was used to identify the differentially expressed genes (DEGs) between control and treatment group. To find out specific genes, DEGs at T0 were compared with DEGs at T1. Scansite was used to find out the binding domain for specific DEGs. Functional enrichment analysis was done by DAVID. RESULTS: Of the genes queried on the Affymetrix Human Genome U133 Plus 2.0 microarray, we identified 263 (T0) and 266 (T1) DEGs compared to the control group. Down-regulation of DEGs related with regulation neuron apoptosis was observed at T1. The most significant function of DEGs at T0 was related with neurological system process. The most specific DEG was FAM125B at T0 and T1 time points. Its common binding domain was SH3. CONCLUSIONS: The protective effect of IPC was associated with altered expression of genes involved in neurological system process and regulation of neuron apoptosis. The dynamic expression of FAM125B can be a supervised marker during the surgery. IPC may be of potential benefit in this and other musculoskeletal conditions.

First Report of Streptomyces galilaeus Associated with Common Scab in China.

During a survey of potato scab pathogens in China from 2003 to 2012, a new pathogen was found in Shanxi and Neimenggu provinces. The incidence was approximately 20% of all recovered strains. The lesions caused by the pathogen were slightly raised and similar to those caused by Streptomyces scabies (3). Lesions were excised (approximately 10 mm3) from 40 infected tubers, surface-disinfested with 0.3% NaOCl for 30 s, rinsed in sterile water three times, cut into 5 mm3, then sliced into 1-mm pieces, and plated on water agar amended with ampicillin (50 µg/ml). Plates were incubated at 28°C in the dark for 4 days. The spores of Streptomyces sp. strains growing from the tuber pieces were collected from single bacterial colonies and cultured on oatmeal agar. To fulfill Koch's postulates, one strain, CPS-2, was grown at 28°C for 10 days and the spores were washed from the plates as inoculum. One hundred milliliters of inoculum (1 × 105 CFU/ml) was mixed with autoclaved soil and vermiculite (1:1) in each pot (15 cm in diameter). Cut tubers were planted in the pots (potato cv. Favorita, one plant per pot, five replicates) and grown under greenhouse conditions (22 ± 5°C). Typical common scab symptoms consisting of small, brown, raised lesions developed on potato tubers 12 weeks after planting. The same strain was re-isolated from the lesions of the new scabby tubers. Non-inoculated plants, treated as described above, but without strain CPS-2, remained healthy. The CPS-2 strain was identified based on morphological and physiological characterization and 16S rDNA sequence. On yeast-malt extract agar, the test strain produced grayish-white aerial hypha, reddish brown substrate mycelium and pigments, and loose spiral spore chains. Spores were smooth and were 0.8 to 0.9 × 1.1 to 1.2 µm in size (diameter and length). The ability of the strain to use single sources of carbon and nitrogen was verified according to the International Streptomyces project (4). The strain grew in media supplemented with L-arabinose, D-fructose, D-glucose, rhamnose, raffinose, meso-inositol, sucrose, and D-xylose, but not D-mannitol. It used L-hydroxyproline, L-methionine, and L-histidine, and produced melanin on tyrosine and peptone yeast extract agar. The strain did not grow at a pH less than 5.0 and was sensitive to streptomycin (20 µg/ml), phenol (0.1%), and crystal violet (0.5 µg/mL), but not to penicillin (10 IU/ml). The strain also produced hydrogen sulfide. The biological characteristics of strain CPS-2 were in accord with Streptomyces galilaeus. CPS-2 produced thaxtomin A in oatmeal liquid medium and the txt AB gene fragment was successfully amplified using specific primers (2). The 16S rDNA sequence of CPS-2 was amplified by PCR with primers 16S1-F: 5'-CATTCACGGAGAGTTTGATCC-3' and 16S1-R: 5'-AGAAAGGAGGTGATCCAGCC-3' (1) and sequenced. A BLAST search of the 16S rDNA sequence for CPS-2 was conducted using the NCBI GenBank database, resulting in 99.8% similarity to S. galilaeus (NR_040857). The 16S rDNA sequence for CPS-2 (1,388 bp) was deposited in GenBank (AY621378). To our knowledge, this is the first report of S. galilaeus causing common scab of potato in China. References: (1) R. A. Bukhalid et al. Appl. Environ. Microbiol. 68:738, 2002. (2) R. Flores-González et al. Plant Pathol. 57:162, 2008. (3) D. H. Lambert and R. Loria. Int. J. Syst. Bacteriol. 39:387, 1989. (4) E. B. Shirling and D. Gottlieb. Int. J. Syst. Bacteriol. 16:313, 1966.

Highly efficient phosphorescent organic light-emitting diode with a nanometer-thick Ni silicide/polycrystalline p-Si composite anode.

A phosphorescent organic light-emitting diode (PhOLED) with a nanometer-thick (approximately 10 nm) Ni silicide/ polycrystalline p-Si composite anode is reported. The structure of the PhOLED is Al mirror/ glass substrate / Si isolation layer / Ni silicide / polycrystalline p-Si/ V(2)O(5)/ NPB/ CBP: (ppy)(2)Ir(acac)/ Bphen/ Bphen: Cs(2)CO(3)/ Sm/ Au/ BCP. In the composite anode, the Ni-induced polycrystalline p-Si layer injects holes into the V(2)O(5)/ NPB, and the Ni silicide layer reduces the sheet resistance of the composite anode and thus the series resistance of the PhOLED. By adopting various measures for specially optimizing the thickness of the Ni layer, which induces Si crystallization and forms a Ni silicide layer of appropriate thickness, the highest external quantum efficiency and power conversion efficiency have been raised to 26% and 11%, respectively.

Anti-ADDL antibodies differentially block oligomer binding to hippocampal neurons.

Abeta-derived diffusible ligands (ADDLs) are abundant in AD brain, bind to hippocampal neurons and induce deficits in rodent cognition. To further investigate ADDL binding to neurons and identify antibodies that block this association, a panel of anti-Abeta and anti-ADDL antibodies was characterized for their ability to immuno-detect neuronally bound ADDLs and attenuate the binding of ADDLs to neurons. The results showed that anti-Abeta and anti-ADDL antibodies were able to abate ADDLs binding to hippocampal neurons, but to different degrees. Quantitative assessment of binding showed that one antibody, ACU-954 was markedly more effective at blocking ADDL binding than other antibodies assessed. ACU-954 was also found to block ADDL binding to hippocampal slice cultures, attenuate the ADDL-induced loss of dendritic spines and detect "natural ADDLs" in human AD tissue. These results demonstrated that antibodies that bind to and block ADDL binding to neurons can be identified, although their efficacy is conformationally specific since it is not readily apparent or predictable based on the core linear epitope or affinity for monomeric Abeta.

Creutzfeldt-Jakob disease in a Chinese patient with a novel seven extra-repeat insertion in PRNP.

Familial transmissible spongiform encephalopathies comprise about 14% of all cases of transmissible spongiform encephalopathy in humans. We report on a patient with a definite diagnosis of familial Creutzfeldt-Jakob disease with an insertional mutation consisting of seven extra octapeptide repeats between codons 51 and 91 in the PRNP gene, associated with a genotype homozygotic for methionine at codon 129 and a novel coding change of the inserted octapeptide region.

Combination of passivated Si anode with phosphor doped organic to realize highly efficient Si-based electroluminescence.

Silicon light source plays a key role in silicon optoelectronics, but its realization is an extremely challenging task. Although there are longterm intensive efforts to this topic, the power conversion efficiency (PCE) of the silicon-based electroluminescence is still no more than 1%. In this present report, a highly efficient silicon light source has been achieved. The device structure is p-Si (5 Omegacm)/ SiO2(approximately 2 nm)/ NPB / CBP: (ppy)(2)Ir(acac) / Bphen /Bphen: Cs2CO3 / Sm / Au. The SiO2 passivated Si is the anode having a suitably high hole-injection ability, and CBP: (ppy)(2)Ir(acac) is a highly efficient phosphor doped organic material. The device turn-on voltage is 3.2 V. The maximum luminance efficiency and maximum luminous power efficiency reach 69 cd/A and 62 lm/W, respectively, corresponding to a maximum PCE of 12% and an external quantum efficiency of 17%.

First Report of Potato Scab Caused by Streptomyces turgidiscabies in China.

Potato scab, caused by several plant pathogenic Streptomyces species, is known to occur in potato-planting areas worldwide. Symptoms of disease on potato tubers are shallow, raised, or pitted corky lesions (2). In 1998, Streptomyces turgidiscabies was reported as a new potato scab pathogen from Hokkaido, Japan (3). Potato scab has been observed in many potato-cultivation areas in China and incidence of the disease was approximately 6 to 10% in some fields in 2006 (in our survey). To investigate the casual agent of scab disease, isolations were made from scabby potato tubers collected from different areas using oatmeal agar. Identification of an isolate from Shaanxi Province was based on morphological and physiological characterization followed by 16S rRNA confirmation. Characteristics were gray, aerial hypha, rectiflexuous spore chains, a smooth spore surface, and spores that were 0.5 to 0.7 × 1.0 to 1.2 µm. The strain did not produce melanin on tyrosine-peptone-yeast extract agar media, did not produce diffusible pigments, used all the International Streptomyces Project (ISP) sugars (4) as single carbon sources, used l-hydroxyproline, l-tyrosine, and l-histidine as single nitrogen sources but not l-methionine, grew at pH 4.5, was susceptible to streptomycin (20 µg ml-1), phenol (0.1%), and penicillin (10 IU ml-1) but not to crystal violet (0.5 µg ml-1), and produced H2S. The identification was confirmed by comparison of its 16S rRNA sequence with the GenBank database using the BLAST program. The 16S rRNA sequence was amplified by PCR with primers S1: 5'-CATTCACGGAGAGTTTGATCC-3' and S2: 5'-AGAAAGGAGGTGATCCAGCC-3' and sequenced. BLASTn analysis of the sequence obtained showed the highest similarity (99.9%) with S. turgidiscabies type strain ATCC 700248 (GenBank Accession No. AB026221). The sequence was submitted to GenBank (Accession No. AM889495). Pathogenicity of the strain was tested in the greenhouse on potato tubers of cv. Favorita grown in pots (one plant per pot, three replicates). One hundred milliliters of inoculum (1 × 105 CFU ml-1) of the strain was mixed with sterile soil and vermiculite (1:1) in each pot. Potato plants were grown at 25°C and the soil was allowed to dry between waterings. The immature potato tubers were used to evaluate scab symptoms 10 weeks after planting. All tubers inoculated with the pathogen developed typical common scab symptoms consisting of erumpent, brown, corky lesions, which is different from the symptoms caused by S. reticuliscabiei (1). The noninoculated control tubers did not show scab symptoms. S. turgidiscabies was reisolated from lesions of diseased immature tubers. The pathogenicity test indicates that S. turgidiscabies caused scab disease on potato tubers. To our knowledge, this is the first report of S. turgidiscabies causing potato scab disease in China. References: (1) K. Bouchek-Mechiche et al. Int. J. Syst. Evol. Microbiol. 56:2771, 2006. (2) R. Loria et al. Plant Dis. 81:836, 1997. (3) K. Miyajima et al. Int. J. Syst. Bacteriol. 48:495, 1998. (4) E. B. Shirling and D. Gottlieb. Int. J. Syst. Bacteriol. 16:313, 1966.

Ethanol blocks both basic fibroblast growth factor- and carbachol-mediated neuroepithelial cell expansion with differential effects on carbachol-activated signaling pathways.

We have expanded neuroepithelial cells dissociated from the embryonic rat telencephalon in serum-free defined medium containing basic fibroblast growth factor (bFGF) in order to generate a model neuroepithelium to study the interaction of ethanol with both growth factor- and transmitter-stimulated proliferation. Ethanol blocked proliferation stimulated by bFGF and by carbachol, an agonist at muscarinic acetylcholine receptors, in a dose-dependent manner. In addition, ethanol attenuated autonomous expansion of neuroepithelial cells occurring following withdrawal of bFGF. The latter effect was associated with an increase in the number of apoptotic cells identified by terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling labeling. We studied the effects of ethanol on carbachol-stimulated signaling pathways critical to its proliferative effects. Ethanol significantly reduced carbachol-stimulated Ca(2+) signaling, as well as Erk1/Erk2, Akt and cyclic AMP-response element-binding phosphorylations in a dose-dependent manner. Comparison of the potency of ethanol in attenuating carbachol-stimulated proliferation and signal transduction showed that mitogen-activated protein kinase phosphorylation was less sensitive to ethanol than the other parameters. The results indicate that ethanol's suppression of proliferation induced by carbachol in this model neuroepithelium likely involves multiple signaling pathways. These effects in vitro may help to explain the devastating effects of prenatal ethanol exposure in vivo, which contribute to the fetal alcohol syndrome.

L-dopa upregulates the expression and activities of methionine adenosyl transferase and catechol-O-methyltransferase.

High nonphysiological doses of l-dopa are administered to Parkinson's disease (PD) patients, to replenish the depleted dopamine (DA). A large portion of the administered L-dopa and the newly formed DA undergoes methylation by reacting with S-adenosyl-L-methionine (SAM). In the process SAM, as well as L-dopa and DA, is utilized and great demands are placed on the transmethylation system. In this study we investigated whether L-dopa increases the transmethylation process by inducing methionine adenosyl transferase (MAT), the enzyme that produces SAM, and catechol-O-methyl transferase (COMT), the enzyme that transfers the methyl group from SAM to L-dopa and DA. Swiss Webster mice were injected with L-dopa, four times/day, for 1 to 16 days. Brain DA, 3-O-methyldopa (3-OMD), SAM, S-adenosylhomocysteine (SAH), MAT, and COMT were measured following a 24-h withdrawal period. An increase of 264% of brain DA occurred at days 2 and 3 after which it tapered to about 164% of control. The brain level of 3-OMD increased to 870% of the control. SAM was increased by 44% after the sixth day and SAH level was about double after the second day. After day 3, MAT activity was increased by about 35%. Western blot analysis showed that MAT is more clearly characterized in 10% mercaptoethanol reducing buffer in which 31.5-, 38- (beta), and 48-kDa (alpha1/alpha2) subunits were distinctly revealed. The induction of the 38-kDa and, more prominently, the 48-kDa subunits of MAT and the potential transactivator proteins of MAT, c-Jun/AP-1, was evident by day 6. The 31.5-kDa subunit was downregulated. COMT was detected as 24.7-, 30-, and 47.5-kDa bands in the brain, consistent with the membrane-bound COMT I (MB-COMT) and the dimeric COMT II. The 24.7- and the 30-kDa MB-COMT bands were induced in the brain by day 6 and peaked on day 9. The highlight of the study is the fact that L-dopa induces the enzymes MAT and COMT. In addition, the downturn in brain DA after the sixth day coincides with the increase in SAM and the 48-kDa MAT protein. Thus, during PD treatment with L-dopa the induction of MAT and COMT is likely to occur and in turn increase the methylation and reduction of L-dopa and DA that may help cause the tolerance or the wearing-off effect developed to L-dopa.

Role of insulin and insulin receptor in learning and memory.

As one of the most extensively studied protein hormones, insulin and its receptor have been known to play key roles in a variety of important biological functions. Until recent years, the functions of insulin and insulin receptor (IR) in the central nervous system (CNS) have largely remained unclear. IR is abundantly expressed in several specific brain regions that govern fundamental behaviors such as food intake, reproduction and high cognition. The IR from the periphery and CNS exhibit differences in both structure and function. In addition to that from the peripheral system, locally synthesized insulin in the brain has also been identified. Accumulated evidence has demonstrated that insulin/IR plays important roles in associative learning, as suggested by results from both interventive and correlative studies. Interruption of insulin production and IR activity causes deficits in learning and memory formation. Abnormal insulin/IR levels and activities are seen in Alzheimer's dementia, whereas administration of insulin significantly improves the cognitive performance of these patients. The synaptic bases for the action of insulin/IR include modifying neurotransmitter release processes at various types of presynaptic terminals and modulating the activities of both excitatory and inhibitory postsynaptic receptors such as NMDA and GABA receptors, respectively. At the molecular level, insulin/IR participates in regulation of learning and memory via activation of specific signaling pathways, one of which is shown to be associated with the formation of long-term memory and is composed of intracellular molecules including the shc, Grb-r/SOS, Ras/Raf, and MEK/MAP kinases. Cross-talk with another IR pathway involving IRS1, PI3 kinase, and protein kinase C, as well as with the non-receptor tyrosine kinase pp60c-src, may also be associated with memory processing.

Theta rhythm of hippocampal CA1 neuron activity: gating by GABAergic synaptic depolarization.

Information processing and memory consolidation during exploratory behavior require synchronized activity known as hippocampal theta (theta) rhythm. While it is well established that the theta activity depends on cholinergic inputs from the medial septum/vertical limb of the diagonal band nucleus (MS/DBv) and theta discharges of GABAergic interneurons, and can be induced with cholinergic receptor agonists, it is not clear how the increased excitation of pyramidal cells could occur with increased discharges of GABAergic interneurons during theta waves. Here, we show that the characteristic theta activity in adult rat hippocampal CA1 pyramidal cells is associated with GABAergic postsynaptic depolarization and a shift of the reversal potential from Cl(-) toward HCO(3)(-) (whose ionic gradient is regulated by carbonic anhydrase). The theta activity was abolished by GABA(A) receptor antagonists and carbonic anhydrase inhibitors, but largely unaffected by blocking glutamate receptors. Carbonic anhydrase inhibition also impaired spatial learning in a water maze without affecting other sensory/locomotor behaviors. Thus HCO(3)(-)-mediated signaling, as regulated by carbonic anhydrase, through reversed polarity of GABAergic postsynaptic responses is implicated in both theta and memory consolidation in rat spatial maze learning. We suggest that this mechanism may be important for the phase forward shift of the place cell discharges for each theta cycle during the animal's traversal of the place field for that cell.

Calexcitin interaction with neuronal ryanodine receptors.

Calexcitin (CE), a Ca2+- and GTP-binding protein, which is phosphorylated during memory consolidation, is shown here to co-purify with ryanodine receptors (RyRs) and bind to RyRs in a calcium-dependent manner. Nanomolar concentrations of CE released up to 46% of the 45Ca label from microsomes preloaded with 45CaCl2. This release was Ca2+-dependent and was blocked by antibodies against the RyR or CE, by the RyR inhibitor dantrolene, and by a seven-amino-acid peptide fragment corresponding to positions 4689-4697 of the RyR, but not by heparin, an Ins(1,4,5)P3-receptor antagonist. Anti-CE antibodies, in the absence of added CE, also blocked Ca2+ release elicited by ryanodine, suggesting that the CE and ryanodine binding sites were in relative proximity. Calcium imaging with bis-fura-2 after loading CE into hippocampal CA1 pyramidal cells in hippocampal slices revealed slow, local calcium transients independent of membrane depolarization. Calexcitin also released Ca2+ from liposomes into which purified RyR had been incorporated, indicating that CE binding can be a proximate cause of Ca2+ release. These results indicated that CE bound to RyRs and suggest that CE may be an endogenous modulator of the neuronal RyR.

Peptidyl-prolyl-cis/trans-isomerase activity may be necessary for memory formation.

At present, evidence for a plethora of physiological roles for the different classes of peptidyl-prolyl-cis/trans-isomerases (PPIases, EC 5.2.1.8) is emerging. Cyclosporin A (CyA) has been previously reported to disrupt memory formation in a temporally specific manner, when administered intracranially to day-old chicks trained on a single-trial, passive-avoidance task [Bennett, P.C., Zhao, W., Lawen, A. and Ng, K.T. (1996) Brain Res. 730, 107-1171. CyA is known to inhibit both the PPIase activity of cyclophilin and, indirectly, the protein phosphatase activity of calcineurin. Therefore to begin to distinguish between these two functions we studied the effects on memory formation of three non-immunosuppressive CyA analogues, in order to study the involvement of cyclophilins. These drugs retain the capacity to bind to and inhibit the PPIase activity of cyclophilin, but do not bind in the complex with cyclophilin to calcineurin and, therefore, do not inhibit its phosphatase activity. All three drugs exert effects on memory formation comparable to those induced by CyA, significantly inhibiting memory formation when injected intracranially (50 fmol per hemisphere) immediately following training. Brain extracts from chicks treated with [MeVal4]CyA show a strong inhibition of cyclophilin activity. These data show a requirement for the PPIase activity of a cyclophilin for successful memory formation and constitute the first set of data establishing a physiological role for a cyclophilin.

Cell cycle-associated accumulation of tissue inhibitor of metalloproteinases-1 (TIMP-1) in the nuclei of human gingival fibroblasts.

We first confirmed an earlier immunohistochemical study showing that immunoreactive TIMP-1-like protein accumulated in the nuclei of human gingival fibroblasts (Gin-1 cells), reaching a maximum in the S phase of the cell cycle (Li, H., Nishio, K., Yamashita, K., Hayakawa, T. and Hoshino, T. (1995). Nagoya J. Med. Sci. 58, 133-142). Then we isolated this protein from a nuclear extract of Gin-1 cells and demonstrated it to be identical to human recombinant TIMP-1 by western blotting, by a sandwich enzyme immunoassay for TIMP-1 and by an assay for matrix metalloproteinase inhibition. The amount of TIMP-1 in the cytosolic fraction of quiescent Gin-1 cells after stimulation by fetal calf serum increased continuously for 48 hours, whereas that in the nuclear extract showed a maximum at 24 hours (S phase) and significantly decreased thereafter. Gin-1 cells expressed mRNAs for both TIMP-2 and TIMP-3 together with mRNA for TIMP-1. However, neither TIMP-2 nor TIMP-3 proteins seemed to accumulate in the nuclei of Gin-1 cells. These facts strongly suggest that TIMP-1 accumulates specifically in the nuclei of Gin-1 cells in a cell cycle-dependent manner.

Chicks injected with antisera to either S-100 alpha or S-100 beta protein develop amnesia for a passive avoidance task.

The cellular expression of S-100 beta protein is upregulated in Alzheimer's disease and in Down's syndrome, and this protein has been implicated in memory-related processes in laboratory animals. However, the possibility that the alpha subunit of S-100 is also involved in memory has not yet been examined. In the present study, day-old black Australorp white Leghorn cockerel chicks (Gallus domesticus) received injections of monoclonal antisera to S-100 alpha (1:50) or S-100 beta (1:500) into each hemisphere immediately after training on a one-trial passive avoidance task. The chicks displayed significantly lower retention levels than control birds that had been injected with antisera to carbonic anhydrase, or with saline (p < .01). S-100 alpha antisera had an amnestic effect when injected between 0 and 20 min after training, with memory deficits occurring from 30 min post-learning, at the point of transition between the A and the B phases of the Gibbs-Ng intermediate memory stage. By contrast, the S-100 beta antisera needed to be injected either 5 min before or immediately after training and produced amnesia 10 min earlier, at the start of the A phase of the intermediate memory stage. We conclude that the two subunits of the S-100 protein are required at different points in the sequence of events leading to the consolidation of passive avoidance memory.